This document discusses embryo culture and micromanipulation techniques used in assisted reproductive technologies. It describes how embryo culture involves allowing embryos to grow artificially before implantation and involves different culture media and techniques like autologous endometrial coculture. It also explains micromanipulation techniques like intracytoplasmic sperm injection, preimplantation genetic diagnosis, and assisted hatching which involve manipulating embryos at a microscopic level.
Embryo transfer is a step in assisted reproduction where embryos are placed in the uterus to establish a pregnancy. There are two types of embryo transfer - fresh and frozen. For frozen transfers, the uterus is prepared with estrogen and progesterone to make it receptive. During the procedure, a catheter is used to insert embryos through the cervix under ultrasound guidance. Patients then wait two weeks before a blood test checks for human chorionic gonadotropin, which would confirm a pregnancy.
Micromanipulation refers to procedures performed using microscopic instruments to manipulate eggs, sperm, and embryos. This includes techniques like partial zone dissection, subzonal insemination, intracytoplasmic sperm injection, somatic cell nuclear transfer, and gene transfer through microinjection. These techniques are used to increase fertilization and pregnancy rates, overcome male infertility, produce clones and chimeras, and enable genetic modification. The first successful transfers of sheep, pig, and cattle embryos occurred in the 1950s, with continuing advances including IVF births in the 1980s and the first animal clone in 1996. However, manipulating animals also raises ethical concerns regarding animal welfare and impacts on the environment.
Cryopreservation allows for the long-term storage of biological tissues like sperm and embryos at sub-zero temperatures, typically in liquid nitrogen. For sperm, the process involves collecting semen samples and freezing them using methods like slow freezing or rapid freezing. Frozen sperm can later be thawed and used in assisted reproduction. Embryo cryopreservation collects embryos after fertilization and selection of high-quality embryos, then freezes and stores the embryos for future use through controlled-rate freezing and thawing processes. Both techniques effectively stop biological activity at low temperatures and allow for indefinite storage of reproductive cells and tissues.
This document discusses effective protocols for superovulation when undergoing IVF treatment. It compares different ovarian stimulation protocols including long and short protocols using gonadotropin-releasing hormone (GnRH) agonists or antagonists. It also examines the use of human menopausal gonadotropin (hMG) versus recombinant follicle-stimulating hormone (r-FSH), as well as adding luteinizing hormone (LH) to stimulation. Key factors discussed include number of eggs retrieved, egg and embryo quality, risk of ovarian hyperstimulation syndrome, and pregnancy rates. The document provides guidance on optimizing protocols based on patient characteristics and treatment goals.
The embryo transfer technique is the final and most crucial step of the IVF cycle. It requires close collaboration between the clinician and embryologist. While around 80% of embryos typically reach the transfer stage, the pregnancy rate remains low due to factors such as poor embryo quality and technique. The success of embryo transfer depends on proper preparation, including evaluating the cervico-uterine axis, clearing mucus from the cervix, and using ultrasound guidance. The embryo must be placed in the optimal location of the uterine cavity to maximize implantation chances. Attention to factors such as catheter type, duration of embryo loading, and prevention of uterine contractions can significantly improve clinical pregnancy outcomes.
This document summarizes cryopreservation techniques for sperm banking and artificial insemination. It discusses how cryopreservation involves freezing biological material like sperm at ultra-low temperatures to preserve it using cryoprotectants like glycerol. Sperm banking involves collecting and storing frozen semen for future artificial insemination procedures to aid fertility. Artificial insemination is then described as the process of using a catheter to inject preserved sperm into the female's reproductive tract to facilitate conception.
This document provides an overview of embryo transfer (ET) in cattle. It describes the ET process, which involves removing embryos from a genetically superior donor cow and transferring them to recipient cows. The goal of ET is to efficiently produce genetically superior offspring. The document outlines the steps of synchronizing donor and recipient cows, flushing embryos from the donor, examining and transferring high quality embryos to recipients. It notes that ET is an expensive but effective way to introduce superior genetics into a herd within a short time period.
The document discusses the Multiple Ovulation Embryo Transfer (MOET) technique used in dairy farming. MOET involves controlling the estrus cycles of donor and recipient cows, superovulating donor cows to produce multiple eggs, artificially inseminating donors, collecting embryos 7-8 days later, and transferring fresh or frozen embryos into recipients. This allows for increasing the herd size more quickly than traditional breeding since one donor cow can produce multiple embryos that can be implanted into various recipients.
Embryo transfer is a step in assisted reproduction where embryos are placed in the uterus to establish a pregnancy. There are two types of embryo transfer - fresh and frozen. For frozen transfers, the uterus is prepared with estrogen and progesterone to make it receptive. During the procedure, a catheter is used to insert embryos through the cervix under ultrasound guidance. Patients then wait two weeks before a blood test checks for human chorionic gonadotropin, which would confirm a pregnancy.
Micromanipulation refers to procedures performed using microscopic instruments to manipulate eggs, sperm, and embryos. This includes techniques like partial zone dissection, subzonal insemination, intracytoplasmic sperm injection, somatic cell nuclear transfer, and gene transfer through microinjection. These techniques are used to increase fertilization and pregnancy rates, overcome male infertility, produce clones and chimeras, and enable genetic modification. The first successful transfers of sheep, pig, and cattle embryos occurred in the 1950s, with continuing advances including IVF births in the 1980s and the first animal clone in 1996. However, manipulating animals also raises ethical concerns regarding animal welfare and impacts on the environment.
Cryopreservation allows for the long-term storage of biological tissues like sperm and embryos at sub-zero temperatures, typically in liquid nitrogen. For sperm, the process involves collecting semen samples and freezing them using methods like slow freezing or rapid freezing. Frozen sperm can later be thawed and used in assisted reproduction. Embryo cryopreservation collects embryos after fertilization and selection of high-quality embryos, then freezes and stores the embryos for future use through controlled-rate freezing and thawing processes. Both techniques effectively stop biological activity at low temperatures and allow for indefinite storage of reproductive cells and tissues.
This document discusses effective protocols for superovulation when undergoing IVF treatment. It compares different ovarian stimulation protocols including long and short protocols using gonadotropin-releasing hormone (GnRH) agonists or antagonists. It also examines the use of human menopausal gonadotropin (hMG) versus recombinant follicle-stimulating hormone (r-FSH), as well as adding luteinizing hormone (LH) to stimulation. Key factors discussed include number of eggs retrieved, egg and embryo quality, risk of ovarian hyperstimulation syndrome, and pregnancy rates. The document provides guidance on optimizing protocols based on patient characteristics and treatment goals.
The embryo transfer technique is the final and most crucial step of the IVF cycle. It requires close collaboration between the clinician and embryologist. While around 80% of embryos typically reach the transfer stage, the pregnancy rate remains low due to factors such as poor embryo quality and technique. The success of embryo transfer depends on proper preparation, including evaluating the cervico-uterine axis, clearing mucus from the cervix, and using ultrasound guidance. The embryo must be placed in the optimal location of the uterine cavity to maximize implantation chances. Attention to factors such as catheter type, duration of embryo loading, and prevention of uterine contractions can significantly improve clinical pregnancy outcomes.
This document summarizes cryopreservation techniques for sperm banking and artificial insemination. It discusses how cryopreservation involves freezing biological material like sperm at ultra-low temperatures to preserve it using cryoprotectants like glycerol. Sperm banking involves collecting and storing frozen semen for future artificial insemination procedures to aid fertility. Artificial insemination is then described as the process of using a catheter to inject preserved sperm into the female's reproductive tract to facilitate conception.
This document provides an overview of embryo transfer (ET) in cattle. It describes the ET process, which involves removing embryos from a genetically superior donor cow and transferring them to recipient cows. The goal of ET is to efficiently produce genetically superior offspring. The document outlines the steps of synchronizing donor and recipient cows, flushing embryos from the donor, examining and transferring high quality embryos to recipients. It notes that ET is an expensive but effective way to introduce superior genetics into a herd within a short time period.
The document discusses the Multiple Ovulation Embryo Transfer (MOET) technique used in dairy farming. MOET involves controlling the estrus cycles of donor and recipient cows, superovulating donor cows to produce multiple eggs, artificially inseminating donors, collecting embryos 7-8 days later, and transferring fresh or frozen embryos into recipients. This allows for increasing the herd size more quickly than traditional breeding since one donor cow can produce multiple embryos that can be implanted into various recipients.
Embryo culture involves growing embryos outside the body in an artificial environment before transferring them to the uterus. Embryos can be cultured for 2-5 days in specialized incubators that carefully control temperature, pH, gas levels and humidity. Extended culture to the blastocyst stage at 5 days allows more selection of healthy embryos and mimics natural implantation timing. A variety of culture media are used optimized for fertilization, early cleavage and blastocyst development stages.
This document discusses sperm cryopreservation, including the aims, techniques, factors affecting results, and future issues. The key points are:
- Sperm cryopreservation preserves sperm cells at sub-zero temperatures for future use, such as for fertility treatments. Slow freezing and rapid freezing are two common techniques.
- Factors like cryoprotectants, cooling/thawing rates, and semen quality can impact sperm survival after thawing. Semen preparation before freezing may improve outcomes.
- While some studies found cryopreservation does not affect reproductive success rates with ICSI, its effects on sperm DNA integrity are still unclear and require more research. Proper cryopreservation protocols aim to minimize DNA damage
Embryo transfer is a technique where embryos are collected from donor females and transferred to recipient females. The key steps are superovulation of the donor, artificial insemination, embryo collection 5-8 days later, grading the embryos, and transferring high quality embryos into a synchronized recipient. This allows one superior donor to produce many offspring, accelerating genetic improvement. Embryo transfer has advantages like faster breeding, disease control and conservation, but requires synchronization of donors and recipients.
Human spermatozoa can tolerate a range of temperature. They are not very sensitive to damage caused by cooling possibly because of high membrane fluidity which is used as a technique to preserve spermatozoa in adverse conditions. cryopreservation technology has been a boon in every aspect of infertility & ART practice.
The document discusses embryo transfer, which is a process where an embryo is collected from a donor female and transferred to a recipient female to complete its development. Embryo transfer allows a genetically superior female to produce more offspring than through natural reproduction. Key aspects discussed include selecting donor females, inducing superovulation in donors to release multiple eggs, inseminating donors, non-surgical and surgical embryo recovery methods, evaluating and storing embryos, and transferring embryos into recipient females through non-surgical or surgical methods.
This document summarizes key concepts in vitrification and cryobiology. It discusses the components involved in vitrification including cryoprotectants like sucrose, ethylene glycol, DMSO, and propylene glycol. It outlines the steps in vitrification - adding cryoprotectants, cooling cells to -196C, warming/thawing, and removing cryoprotectants. Variables that can influence vitrification effectiveness are discussed. Composition of vitrification and warming solutions from different studies are presented. Similar survival rates between vitrification and slow freezing are noted. Open questions around water removal from oocytes during vitrification are raised. The need for long term follow up studies on neonatal outcomes is emphasized.
This document discusses embryo transfer as the final step of assisted reproductive technology where embryos are placed in the female uterus. It describes factors that affect embryo transfer success including implantation capacity, endometrial receptivity, and technique. The document outlines techniques for embryo transfer including assisted hatching, embryo glue, catheter type and loading, ultrasound guidance, and ensuring no blood or mucus blockage. Successful embryo transfer requires a gentle, non-traumatic procedure performed with attention to numerous technical details.
In vitro maturation and In vitro FertilizationAsadullah Babar
Immature eggs are retrieved from the ovary through aspiration or slicing and matured in the laboratory. The eggs are then fertilized in vitro through incubation with sperm for 8-10 hours. Finally, the presumed zygotes are cultured for 9 days to allow embryonic development. This process involves collection, transportation, in vitro maturation of eggs, in vitro fertilization through sperm treatment and incubation, and in vitro culture of the resulting embryos.
This document discusses modern techniques for pre-selecting the sex of embryos, which can be done either through sperm sorting by looking for X or Y chromosomes, or by determining the sex of pre-implantation embryos. There are invasive methods like cytogenetic analysis of chromosomes and DNA probes, and non-invasive methods like detecting the H-Y antigen or differences in growth between male and female embryos. These techniques allow for altering the male to female ratio in farm animals to increase milk and meat production.
In vitro fertilization and embryo transfer in humansHasnahana Chetia
The document discusses infertility treatment techniques like in vitro fertilization (IVF) and embryo transfer. It describes the IVF process which involves collecting eggs and sperm, fertilizing the eggs in vitro, and implanting the resulting embryos into the uterus. Embryo transfer is defined as implanting embryos developed in vitro or from another female's uterus. The success rates of IVF depend on factors like the woman's age and number of eggs collected. IVF has led to the birth of the first "test tube baby" and advances in reproductive technology.
This document provides an overview of intracytoplasmic sperm injection (ICSI). ICSI involves injecting a single sperm directly into a mature egg under a microscope, which differs from conventional in vitro fertilization where fertilization occurs outside the body. ICSI may be recommended for couples experiencing issues like low sperm counts, abnormal sperm, or problems with antibodies. The process involves sperm and egg retrieval followed by cleaning and injecting a sperm into an egg's cytoplasm. Success rates vary depending on patient factors, though ICSI enables fertilization when few sperm are available. Potential risks include genetic issues being passed to offspring if the father's sperm is abnormal.
Embryo freezing involves preserving embryos at sub-zero temperatures, usually before implantation. There are two main methods - controlled rate freezing and vitrification. Controlled rate freezing involves slow cooling and seeding to prevent ice crystal formation, while vitrification solidifies the solution without any ice crystals using high concentrations of cryoprotectants. Both aim to minimize damage to embryos from freezing and thawing. Cryopreservation allows storage of surplus embryos and avoids multiple pregnancies from single retrievals. It also enables disease screening before transfer.
Title "In vitro production of embryos from high performance cows and the development of frozen-thawed embryos after transfer". This presentation was from reviewed journal that published on 2008.
The document discusses embryonic development from an embryologist's perspective. It covers the overall view of embryonic development from conception through harvest and techniques. The source and significance of embryonic development is also examined at a high level.
Micromanipulation refers to microscopic techniques used to manipulate eggs, sperm, and embryos to increase the chances of a successful in vitro fertilization (IVF) cycle. Common micromanipulation techniques include intracytoplasmic sperm injection (ICSI), preimplantation genetic diagnosis (PGD), and assisted hatching. PGD involves testing a single cell from an early embryo for genetic abnormalities, while ICSI involves directly injecting a single sperm into an egg. Assisted hatching involves weakening an embryo's outer shell to improve implantation chances. These micromanipulation techniques have helped address male factor infertility, diagnose genetic conditions, and hold promise for further IVF advancements.
Introduction
Definition
History
Why are the transgenic animals being produced
Transgenic mice
Mice: as model organism
Methods of creation of transgenic mice
knock-out mice
Application of transgenic mice
Conclusion
References
This document discusses assisted reproductive technologies (ART) such as artificial insemination, in vitro fertilization, and intracytoplasmic sperm injection. It describes the various types of ART, including artificial insemination using donor sperm or partner sperm, IVF which involves fertilizing eggs outside the body then transferring embryos, and ICSI which is used when sperm count is low. The document outlines the basic process of IVF and discusses alternatives like GIFT and ZIFT. It also notes ethics concerns and the lack of comprehensive regulation in India.
Embryo transfer in cattle and its applicationsReenaNegi13
Embryo transfer in cattle involves removing embryos from donor cows and transferring them to recipient cows in order to propagate desirable genetic traits. Key steps include superovulating donors to produce multiple embryos, artificially inseminating donors, flushing embryos from donors 6-8 days later, evaluating and cryopreserving embryos, synchronizing recipients, and surgically or non-surgically transferring embryos into recipients. Embryo transfer allows for genetic improvement of cattle herds, planned mating of elite animals, and circumvention of infertility issues.
This document discusses various assisted reproductive techniques (ART) used to treat infertility. It begins by defining ART and infertility. It then describes techniques like artificial insemination (IUI, ICI), in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), gamete intrafallopian transfer (GIFT), zygote intrafallopian transfer (ZIFT), and surrogacy. For each technique, it provides details on the procedure, indications for use, and success rates. It concludes by discussing some risks associated with IVF and ICSI and outlining the two types of gestational surrogacy.
Embryo culture involves growing embryos outside the body in an artificial environment before transferring them to the uterus. Embryos can be cultured for 2-5 days in specialized incubators that carefully control temperature, pH, gas levels and humidity. Extended culture to the blastocyst stage at 5 days allows more selection of healthy embryos and mimics natural implantation timing. A variety of culture media are used optimized for fertilization, early cleavage and blastocyst development stages.
This document discusses sperm cryopreservation, including the aims, techniques, factors affecting results, and future issues. The key points are:
- Sperm cryopreservation preserves sperm cells at sub-zero temperatures for future use, such as for fertility treatments. Slow freezing and rapid freezing are two common techniques.
- Factors like cryoprotectants, cooling/thawing rates, and semen quality can impact sperm survival after thawing. Semen preparation before freezing may improve outcomes.
- While some studies found cryopreservation does not affect reproductive success rates with ICSI, its effects on sperm DNA integrity are still unclear and require more research. Proper cryopreservation protocols aim to minimize DNA damage
Embryo transfer is a technique where embryos are collected from donor females and transferred to recipient females. The key steps are superovulation of the donor, artificial insemination, embryo collection 5-8 days later, grading the embryos, and transferring high quality embryos into a synchronized recipient. This allows one superior donor to produce many offspring, accelerating genetic improvement. Embryo transfer has advantages like faster breeding, disease control and conservation, but requires synchronization of donors and recipients.
Human spermatozoa can tolerate a range of temperature. They are not very sensitive to damage caused by cooling possibly because of high membrane fluidity which is used as a technique to preserve spermatozoa in adverse conditions. cryopreservation technology has been a boon in every aspect of infertility & ART practice.
The document discusses embryo transfer, which is a process where an embryo is collected from a donor female and transferred to a recipient female to complete its development. Embryo transfer allows a genetically superior female to produce more offspring than through natural reproduction. Key aspects discussed include selecting donor females, inducing superovulation in donors to release multiple eggs, inseminating donors, non-surgical and surgical embryo recovery methods, evaluating and storing embryos, and transferring embryos into recipient females through non-surgical or surgical methods.
This document summarizes key concepts in vitrification and cryobiology. It discusses the components involved in vitrification including cryoprotectants like sucrose, ethylene glycol, DMSO, and propylene glycol. It outlines the steps in vitrification - adding cryoprotectants, cooling cells to -196C, warming/thawing, and removing cryoprotectants. Variables that can influence vitrification effectiveness are discussed. Composition of vitrification and warming solutions from different studies are presented. Similar survival rates between vitrification and slow freezing are noted. Open questions around water removal from oocytes during vitrification are raised. The need for long term follow up studies on neonatal outcomes is emphasized.
This document discusses embryo transfer as the final step of assisted reproductive technology where embryos are placed in the female uterus. It describes factors that affect embryo transfer success including implantation capacity, endometrial receptivity, and technique. The document outlines techniques for embryo transfer including assisted hatching, embryo glue, catheter type and loading, ultrasound guidance, and ensuring no blood or mucus blockage. Successful embryo transfer requires a gentle, non-traumatic procedure performed with attention to numerous technical details.
In vitro maturation and In vitro FertilizationAsadullah Babar
Immature eggs are retrieved from the ovary through aspiration or slicing and matured in the laboratory. The eggs are then fertilized in vitro through incubation with sperm for 8-10 hours. Finally, the presumed zygotes are cultured for 9 days to allow embryonic development. This process involves collection, transportation, in vitro maturation of eggs, in vitro fertilization through sperm treatment and incubation, and in vitro culture of the resulting embryos.
This document discusses modern techniques for pre-selecting the sex of embryos, which can be done either through sperm sorting by looking for X or Y chromosomes, or by determining the sex of pre-implantation embryos. There are invasive methods like cytogenetic analysis of chromosomes and DNA probes, and non-invasive methods like detecting the H-Y antigen or differences in growth between male and female embryos. These techniques allow for altering the male to female ratio in farm animals to increase milk and meat production.
In vitro fertilization and embryo transfer in humansHasnahana Chetia
The document discusses infertility treatment techniques like in vitro fertilization (IVF) and embryo transfer. It describes the IVF process which involves collecting eggs and sperm, fertilizing the eggs in vitro, and implanting the resulting embryos into the uterus. Embryo transfer is defined as implanting embryos developed in vitro or from another female's uterus. The success rates of IVF depend on factors like the woman's age and number of eggs collected. IVF has led to the birth of the first "test tube baby" and advances in reproductive technology.
This document provides an overview of intracytoplasmic sperm injection (ICSI). ICSI involves injecting a single sperm directly into a mature egg under a microscope, which differs from conventional in vitro fertilization where fertilization occurs outside the body. ICSI may be recommended for couples experiencing issues like low sperm counts, abnormal sperm, or problems with antibodies. The process involves sperm and egg retrieval followed by cleaning and injecting a sperm into an egg's cytoplasm. Success rates vary depending on patient factors, though ICSI enables fertilization when few sperm are available. Potential risks include genetic issues being passed to offspring if the father's sperm is abnormal.
Embryo freezing involves preserving embryos at sub-zero temperatures, usually before implantation. There are two main methods - controlled rate freezing and vitrification. Controlled rate freezing involves slow cooling and seeding to prevent ice crystal formation, while vitrification solidifies the solution without any ice crystals using high concentrations of cryoprotectants. Both aim to minimize damage to embryos from freezing and thawing. Cryopreservation allows storage of surplus embryos and avoids multiple pregnancies from single retrievals. It also enables disease screening before transfer.
Title "In vitro production of embryos from high performance cows and the development of frozen-thawed embryos after transfer". This presentation was from reviewed journal that published on 2008.
The document discusses embryonic development from an embryologist's perspective. It covers the overall view of embryonic development from conception through harvest and techniques. The source and significance of embryonic development is also examined at a high level.
Micromanipulation refers to microscopic techniques used to manipulate eggs, sperm, and embryos to increase the chances of a successful in vitro fertilization (IVF) cycle. Common micromanipulation techniques include intracytoplasmic sperm injection (ICSI), preimplantation genetic diagnosis (PGD), and assisted hatching. PGD involves testing a single cell from an early embryo for genetic abnormalities, while ICSI involves directly injecting a single sperm into an egg. Assisted hatching involves weakening an embryo's outer shell to improve implantation chances. These micromanipulation techniques have helped address male factor infertility, diagnose genetic conditions, and hold promise for further IVF advancements.
Introduction
Definition
History
Why are the transgenic animals being produced
Transgenic mice
Mice: as model organism
Methods of creation of transgenic mice
knock-out mice
Application of transgenic mice
Conclusion
References
This document discusses assisted reproductive technologies (ART) such as artificial insemination, in vitro fertilization, and intracytoplasmic sperm injection. It describes the various types of ART, including artificial insemination using donor sperm or partner sperm, IVF which involves fertilizing eggs outside the body then transferring embryos, and ICSI which is used when sperm count is low. The document outlines the basic process of IVF and discusses alternatives like GIFT and ZIFT. It also notes ethics concerns and the lack of comprehensive regulation in India.
Embryo transfer in cattle and its applicationsReenaNegi13
Embryo transfer in cattle involves removing embryos from donor cows and transferring them to recipient cows in order to propagate desirable genetic traits. Key steps include superovulating donors to produce multiple embryos, artificially inseminating donors, flushing embryos from donors 6-8 days later, evaluating and cryopreserving embryos, synchronizing recipients, and surgically or non-surgically transferring embryos into recipients. Embryo transfer allows for genetic improvement of cattle herds, planned mating of elite animals, and circumvention of infertility issues.
This document discusses various assisted reproductive techniques (ART) used to treat infertility. It begins by defining ART and infertility. It then describes techniques like artificial insemination (IUI, ICI), in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), gamete intrafallopian transfer (GIFT), zygote intrafallopian transfer (ZIFT), and surrogacy. For each technique, it provides details on the procedure, indications for use, and success rates. It concludes by discussing some risks associated with IVF and ICSI and outlining the two types of gestational surrogacy.
This document discusses various assisted reproductive techniques (ART) used to treat infertility. It begins by defining ART and infertility. It then describes techniques like artificial insemination (IUI, ICI), in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), gamete intrafallopian transfer (GIFT), zygote intrafallopian transfer (ZIFT), and surrogacy. For each technique, it provides details on the procedure, indications for use, and success rates. It concludes by discussing some risks associated with IVF and ICSI, like prematurity, low birth weight, and possible genetic or epigenetic abnormalities in offspring.
The document summarizes the history and process of in vitro fertilization (IVF). It discusses how IVF was developed as a treatment for infertility and involves fertilizing an egg outside of the body. The summary includes:
- The key stages of an IVF cycle including ovarian stimulation, egg retrieval, fertilization in vitro, embryo transfer, and indications for IVF such as tubal disease or male factor infertility.
- Milestones in the history of IVF including the first successful case in 1978 and development of techniques like ICSI.
- Risks and side effects of ovarian stimulation and factors considered for embryo transfer like number based on patient age and prior attempts.
2- Embryo Transfer in farm animal23.pptEssamTharwat9
This document discusses biotechnology in animal reproduction, focusing on embryo transfer. It defines embryo transfer as collecting embryos from donor females and transferring them to recipient females. The key steps are estrus synchronization of donors and recipients, superovulation of donors to produce multiple embryos, artificial insemination of donors, non-surgical or surgical embryo collection, grading and selection of high quality embryos, and transferring embryos into the uterus of synchronized recipients. The goals are to distribute genetics more efficiently and produce offspring from elite females without compromising their productivity.
Embryo transfer techniques allow for obtaining multiple foals from genetically valuable mares. The process involves flushing embryos from a donor mare 7-8 days after insemination and surgically transferring them into a synchronized recipient mare. Key steps include choosing young, fertile donor mares; carefully timing insemination, flushing, and transfer to match the donor and recipient's cycles; and providing post-procedure care to support pregnancy. While embryo transfer is an important tool for horse breeding, challenges remain in techniques for older mares and reducing costs to make the technology more accessible.
EMBRYOSPLITTING IS THE TECHNIQUE WHEREBY AN EARLY STAGE EMBRYO SPLIT INTO TWO OR MORE GENETICALLY IDENTICAL EMBRYOS. THERE ARE CURRENTLY NUMBER OF TECHNIQUES USED TO CREATE CLONED EMBRYOS, SUCH AS SOMATIC CELL NUMCLEAR TRANSFER, THERAPEUTIC CLONING, EMBRYO SPLITTING AND PARTHENIGENESIS.
EMBRYO SPLITTING IS ALSO KNOWN AS ARTIFICIAL TWINS, MAMMALIAN EMBRYOS SPLITING HAS SUCCESFULLY BEEN ESTABLISHED IN FARM ANIMALS.
Infertility is a growing problem caused by factors like delayed childbearing, diseases, pollution, diet, lack of exercise, and prior contraceptive or abortion procedures. In-vitro fertilization (IVF) involves fertilizing an egg outside the body by mixing it with sperm in a laboratory dish. The fertilized egg is then transferred to the uterus. Key steps in IVF include ovarian stimulation to produce multiple eggs, egg retrieval, fertilizing the eggs with sperm in vitro, embryo culture, and embryo transfer. IVF has allowed many infertile couples to conceive and has resulted in over 5 million births worldwide.
EMBRYO CULTURE :-
"The embryo of different developmental stages, formed within the female gametophyte through sexual process, can be isolated aseptically from the bulk of maternal tissues of ovule, seed or capsule and cultured in vitro under aseptic and controlled physical conditions in glass vials containing nutrient solid or liquid medium to grow directly into plantlet."
The document provides details on the various stages of the in vitro fertilization (IVF) process. It discusses (1) ovulation induction and egg retrieval, which involves monitoring the ovaries and using fertility drugs to stimulate egg production before extracting the eggs, (2) fertilization either naturally or through intracytoplasmic sperm injection (ICSI) if needed, (3) embryo culture and selection of the best embryos for transfer, and (4) embryo transfer involving placing embryos back into the uterus. It also covers success rates, additional procedures like assisted hatching and embryo freezing, and alternatives when sperm is unavailable.
Microinsemination involves directly injecting a single sperm into an egg and can help couples with male factor infertility issues undergo in vitro fertilization successfully. It is performed when sperm have poor motility, low density, defects, or cannot penetrate an egg naturally. The process involves preparing sperm, holding an egg in place with a pipette, drawing up a single sperm into a needle, and injecting it into the egg's cytoplasm. This restores normal fertilization rates. While technical issues or lack of success are risks, data does not suggest an increased risk of birth defects, and it may increase pregnancy chances for those with reduced fertilization through standard IVF.
This document discusses the technique of embryo splitting. It describes how embryo splitting involves removing cells from an early stage embryo and placing them in a nutrient solution to develop into genetically identical embryos. There are two main methods: blastomere biopsy, which removes cells from the embryo through an opening in the zona pellucida, and bisection of the embryo, which divides it into two halves. Embryo splitting can provide more embryos for implantation and may help patients with low response to fertility treatments. However, the clones produced are not genetically identical to the parents and the technique has a low success rate.
In vitro Fertilization- IVF is a form of assisted reproductive technology (ART).In this special medical techniques are used to help a woman become pregnant. IVF, coupled with embryo transfer, in humans is aimed to enable couples suffering from certain types of sterility to have children.
Babies developed from this approach are known as "test-tube babies."
The document discusses in vitro fertilization (IVF), a type of assisted reproductive technology used to treat infertility. IVF involves fertilizing an egg outside the body, in vitro, through the mixing of sperm and egg in a laboratory dish, and then transferring the embryo to the uterus. The document outlines the IVF process, including ovarian stimulation, egg retrieval, sperm collection, fertilization in the lab, embryo culture, and embryo transfer. It notes IVF can result in 70-80% of fertilized eggs developing and the first successful IVF birth occurring in 1978. Risks of IVF are also summarized.
This document provides an overview of several assisted reproductive technologies (ART) including in vitro fertilization (IVF), intrauterine insemination (IUI), intracytoplasmic sperm injection (ICSI), gamete intrafallopian transfer (GIFT), and zygote intrafallopian transfer (ZIFT). It describes the basic procedures and steps for IVF including ovarian stimulation, egg retrieval, fertilization, embryo culture, and embryo transfer. It notes some common indications for IVF include tubal disease, endometriosis, ovulatory dysfunction, and male factor infertility. Potential complications of IVF like ovarian hyperstimulation syndrome and multiple pregnancies are also outlined. IUI and ICSI procedures
1. Infertility is defined as the inability to conceive after one year of unprotected intercourse. Around 10-15% of the population experiences infertility.
2. There are several requirements for natural conception, including the production of healthy eggs and sperm, unblocked fallopian tubes, fertilization, and implantation. When these requirements are not met, Assisted Reproductive Technologies (ART) may help.
3. ART includes procedures like in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), frozen embryo transfer, and pre-implantation genetic screening. The goal is to help sperm fertilize eggs or implant embryos when natural conception is not possible. Selecting an experienced ART
This document discusses various aspects of assisted reproductive technology (ART) including in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). It provides information on the causes of infertility, procedures for IVF such as egg retrieval, embryo transfer, and blastocyst culture. The document also discusses who may benefit from IVF and ICSI, including those with male factor infertility issues or previous IVF failures. Other ART procedures mentioned include zygote intrafallopian transfer, gamete intrafallopian transfer, and potential future techniques like creating artificial gametes.
This document discusses various aspects of assisted reproductive technology (ART) including infertility, its causes, treatments, and specific procedures like in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). It provides information on male and female infertility, decreasing male fertility rates, and drugs to treat male infertility. The document also explains the procedures for IVF such as ovulation induction, egg retrieval, fertilization, embryo culture, and embryo transfer. ICSI is introduced as a technique used to treat male factor infertility.
The document discusses the importance of assisted reproductive biotechnology in humans and animals. It notes that about 1 in 6 couples is infertile in the US and look to clinics in India and Asia for help. It also discusses efforts to save endangered species like the Pashmina goat and wild buffalo through cloning techniques. The document then goes on to discuss various causes of male and female infertility and techniques used in assisted reproduction like IVF, ICSI, and embryo transfer.
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2. CONTENT
EMBRYO CULTURE
• What Is An Embryo Culture
• How It Is Performed
• Techniques In Embryo Culture
MICROMANIPULATION
• Introduction
• Widely Used Techniques For Micromanipulation
• Conclusion
3. What Is An Embryo Culture
• Embryo culture is a component of in vitro fertilization where in
resultant embryos are allowed to grow for sometime in an artificial
medium before being inserted into the uterus.
4. HOW IS IT PERFORMED : by 2-ways :-
Artificial culture
Single
culture
Sequential
culture
Autologous
endometrial
coculture
5. ARTIFICIAL EMBRYO CULTURE MEDIA
• It contains glucose, pyruvate and energy providing components.
• Amino-acids, nucleotides, vitamins and cholesterol improve the
performance of embryonic growth and development.
• SINGLE CULTURE : the same culture medium throughout the medium.
• SEQUENTIAL CULTURE : the embryo is sequentially placed in different
medium.
6. AUTOLOGOUS ENDOMETRIAL COCULTURE
• Autologous endometrial coculture is a technique of assisted
reproductive technology . It involves placing a patient’s fertilized eggs
on top of a layer of cells from her own uterine lining, creating a more
natural environment for embryo development and maximizing the
chance for an in vitro fertilization (IVF) pregnancy.
7. HOW COCULTURE IS PERFORMED
• Patient undergoes an endometrial biopsy during which a small piece
of her uterine lining is removed.
• The uterine lining sample is sent to a research lab, where it is treated,
purified and frozen.
• The patient then undergoes a typical IVF cycle and is given
medication to stimulate egg growth in her ovaries.
• The patient’s eggs are retrieved and mixed with the sperm. At this
time, the lab begins thawing and growing her endometrial cells.
8. • Once fertilization is confirmed, the patient’s embryos are placed on
top of her own (and now thawed) endometrial cells.
Over the next two days, the embryos are closely monitored for
growth and development.
The patient’s embryos are transferred into her uterus for
implantation and pregnancy
Coculture can be an effective treatment for patients who have failed
previous IVF cycles or who have poor embryo quality.
10. • OOCYTE WASH BUFFER
• FERTILIZATION MEDIUM
• CLEAVAGE MEDIUM BLASOCYST MEDIUM
• SPERM BUFFER
• SPERM MEDIUM
• BLASTOCYST CULTURE
11. 1. OOCYTE WASH BUFFER
• On the day of egg retrieval (Day 0), this buffer is used for the retrieval
of the eggs from the ovary. Oocyte wash buffer has an ingredient,
which prevents a change in pH when the solution is exposed to air
during the retrieval. The eggs are very susceptible to any minute
changes in the pH of their environment. The eggs are washed in this
buffer and then placed into the next medium for culture.
12. 2. FERTILIZATION MEDIUM
• After the wash at retrieval, the eggs are put into the fertilization
medium. This medium contains a variety of salts, sugars, amino acids,
protein and other nutrients essential for the maintenance of the egg
(and sperm in IVF) during the process of fertilization (IVF and ICSI).
The fertilization medium and all of the other subsequent culture
media, are buffered with the appropriate components in order to
maintain the correct pH of the solution in the embryo incubator.
13. 3. CLEAVAGE MEDIUM
• All of the eggs which undergo normal fertilization are next placed into
cleavage medium, which is formulated specifically to support the
growth requirements of the early cleavage stage embryo. The
cleaving (dividing) embryo is cultured in this medium until Day 3. If
the embryo transfer is scheduled for Day 3, the embryos are
transferred to the uterus in a small amount of this medium.
14. 4. BLASTOCYST
MEDIUM
• Embryos, that are to be cultured until Day 5
or 6, are placed, later on Day 3, into another
medium referred to as blastocyst medium.
The embryos are then maintained in this
medium until embryo transfer on Day 5 or
embryo cryopreservation on Day 5 or 6. This
medium has additional components and/or
different components required by the
embryo in its transition from a cleavage
stage embryo to a blastocyst. If the embryo
transfer is scheduled on Day 5, the embryos
are transferred to the uterus in a small
amount of this medium.
15. 5. SPERM BUFFER
• The sperm buffer is formulated in order to maintain the correct pH
when the solution is exposed to air. This buffer is used during the
preparation of semen samples and solutions for semen samples,
which will be washed and processed outside of the incubator.
16. 6. SPERM MEDIUM
• The sperm medium is similar to the Sperm Buffer except that the
buffer is such that the correct pH of the solution is maintained whilst
in the incubator. This medium is important for the final resuspension
of sperm to be used in IVF because the process of fertilization occurs
inside the incubator.
17. HOW LONG SHOULD AN EMBRYO BE
CULTURED
• Two Days: Embryos that are cultured for two days are generally transferred at the
two or four-cell stage. This type of transfer is beneficial for couples who have a
low number of embryos available for transfer, or who have embryos that are
developing poorly.
• Three Days: Embryos that are cultured for three days are usually transferred at
the six to eight cell stage. Many laboratories prefer to culture embryos until this
stage because it allows for increased monitoring.
• Five Days: Embryos that are cultured for five days are transferred at the
blastocyst stage. Blastocysts consist of 12 to 16 cells and are well on their way to
be ready for implantation into the uterus. Many labs opt to transfer at the
blastocyst stage, particularly if you have had repeated miscarriages or IVF failures.
19. INTRODUCTION
• In 1966, Lin described the technique of micromanipulating and injecting a mouse
egg.
• Subsequently, transgenic animals have been produced by introduction of foreign
genes at the pronuclear stages of fertilized, one-cell zygotes.
• Most of the successes have been with mouse and recently successful production
of transgenic rabbit, pig, sheep and goat have been shown.
• This technique is a powerful tool for studying gene regulation and physiological
functions of gene products in a normal host environment.
• Micromanipulation is the technique whereby sperm, eggs and embryos can be
handled on an inverted microscope stage, performing minute procedures at the
microscopic level via joysticks that hydraulically operate glass microtools.
20. WIDELY USED TECHNIQUES FOR
MICROMANIPULATION
1. Intra cytoplasmic sperm injection (ICSI)
2. Preimplantation genetic diagnosis (PGD)
3. Assisted hatching
21. INTRA CYTOPLASMIC SPERM INJECTION
• The injection of a single sperm into the cytoplasm of the
oocyte, or intracytoplasmic sperm injection (ICSI),
provided a satisfactory solution to the problems of the
assisted fertilization techniques developed earlier.
• In this procedure, a single sperm is first immobilized by
touching the sperm tail with an injection pipette (inner
diameter 5–7 μm). The injection pipette picks up the
immobilized sperm, pierces the ZP and oolemma, and
delivers the sperm inside the oocyte cytoplasm.
• In 1976 using hamsters as a model, Uehara and
Yanagimachi were probably the first to report the injection
of sperm into oocyte cytoplasm (ooplasm).
• It was later attempted on rabbit and human oocytes,
although the first successful human pregnancy was not
reported until 1992 by the Free University of Brussels’
group in Belgium.
22. PREIMPLANTATION GENETIC DIAGNOSIS
What is it?
• Genetic analysis of a single cell from an eight-cell embryo done in conjunction with
in vitro fertilization (IVF) to improve the chances of a “normal” pregnancy.
Why consider PGD in addition to IVF?
1. recurrent miscarriages
2. one child already affected with a genetic disease
3. family history of inherited disease
4. maternal age older than 38
5. prior failure with IVF
6. family “balancing” for sex
23. METHOD OF
PREIMPLANTATION
GENETIC DIAGNOSIS
1. A single cell from the 6-8-cell embryo is removed using a
fine glass needle by puncturing the zona pellucida and
aspirate the cell. - In skilled hands, this generally does not
harm the developing embryo. - Each cell is called a
blastomere.
• Blastomere removal for PGD testing
2. Prepare a metaphase spread of chromosomes to assess
karyotype (number and integrity of each chromosome)
Two types of assessment techniques are common:
a. chromosome “painting” (or FISH) using fluorescent
probes specific for each chromosome.
b. Polymerase chain reaction (PCR) - amplification of DNA
specific to a gene of interest (family history guides choice
of genes)
24. ASSISTED HATCHING
• Assisted hatching is an IVF technique in which the zona is treated
prior to embryo transfer in order to weaken the wall of the embryo
and thus improve the likelihood of successful hatching and embryo
implantation.
• Just prior to embryo transfer, the developing embryo must "hatch”
out of its outer shell (zona pellucida).
• Some embryos seem to have a thicker shell that may decrease their
ability to implant .
• In such cases Advance technique of IVF with assisted hatching is
performed.
25. TECHNIQUES USED IN
ASSISTED HATCHING
1. MECHANICAL HATCHING : In this the embryo cover is slit
open mechanically with the help of a thin long drawn out
glass needle. This is done with the help of a machine
called micromanipulator
2. CHEMICAL HATCHING : Creating a hole in the zona
pellucida of an embryo by using acidic Tyrode's solution .
3. LASER-ASSISTED HATCHING : this technique involves the
creation of a precise gap in the zona pellucida of selected
embryos using a 1.48 micron infrared diode laser.
Contact laser and Non contact laser
• Contact laser : laser radiation has to be delivered directly to
the zona called contact laser
• Non contact laser : laser gamete manipulation has described
in the non-contact mode.
26. CONCLUSION
• Micromanipulation technology has enabled the reproductive biologist to
overcome inefficient steps in mammalian fertilization, the production of chimeric
animals through blastocyst injection with embryonic stem (ES) cells and the
introduction of specific genes into the genomes of domestic and laboratory
animals.
• This technology has also been used for the production of cloned animals and ES
cell lineages from cloned embryos, using nuclear transfer. Moreover,
micromanipulation is also used for microsurgical embryo biopsy to study the basic
developmental biology of embryos during preimplantation development.