Primary cell cultures are derived directly from animal tissues and have a limited lifespan, usually undergoing fewer than 10 divisions. They retain characteristics of the original tissue. Diploid cell strains can undergo 20-50 passages while maintaining the original karyotype. Continuous cell lines are immortalized cell lines that can divide indefinitely, having undergone changes including aneuploidy and loss of differentiation. Common types of cell culture include primary cultures from tissues like monkey kidney, diploid strains from fetal tissues like human lung fibroblasts, and continuous lines derived from tumors.
Cellular coning refers to generation of genetically identical cells from parent cells. This presentation teaches differences between cell coning and molecular cloning and various methods of cell cloning. Sample questions are also provided for your review of concept learned
Introduction
What is cloning?
Why we want to do cloning?
History
Technique of cell cloning
Dolly – the sheep
Species cloned
Why persue animal cloning research?
Conclusion
Introduction
What is cloning?
Why we want to do cloning?
History
Technique of cell cloning
Dolly – the sheep
Species cloned
Why persue animal cloning research?
Conclusion
Introduction
History
Cell culture techniques
Species cloned
Approaches of cell cloning
Monolayer culture- Dilution cloning
Microtitration plate
Suspension culture- Cloning in agar
Cloning in methocel
Isolation of clone
By clonal rings
By suspension clone
Application of cell cloning
Conclusion
Reference
Cellular coning refers to generation of genetically identical cells from parent cells. This presentation teaches differences between cell coning and molecular cloning and various methods of cell cloning. Sample questions are also provided for your review of concept learned
Introduction
What is cloning?
Why we want to do cloning?
History
Technique of cell cloning
Dolly – the sheep
Species cloned
Why persue animal cloning research?
Conclusion
Introduction
What is cloning?
Why we want to do cloning?
History
Technique of cell cloning
Dolly – the sheep
Species cloned
Why persue animal cloning research?
Conclusion
Introduction
History
Cell culture techniques
Species cloned
Approaches of cell cloning
Monolayer culture- Dilution cloning
Microtitration plate
Suspension culture- Cloning in agar
Cloning in methocel
Isolation of clone
By clonal rings
By suspension clone
Application of cell cloning
Conclusion
Reference
Primary and established cell line cultureKAUSHAL SAHU
Introduction
Primary Culture
Steps of Primary Culture
Isolation Of Tissue
Dissection And Disaggregation
Types Of Primary Culture
Primary Explants Culture
Enzymatic Disaggregation
Mechanical Disaggregation
Cell Line( Finite & Continuous)
Naming A Cell Line
Choosing A Cell Line
Maintenance Of Cell Line
Conclusion
Reference
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
SYNCHRONOUS CULTURES CAN BE OBTAINED IN SEVERAL WAYS:
Physical fractionation .
Chemical appro ach
CENTRIFUGAL ELUTRIATION
Inhibition of DNA synthesis
Nutritional deprivation
SYNCHRONIZATION AT LOW TEMPERATURE
CELLULAR TOTIPOTENCY
SOME HIGHLIGHTS OF CELL SYNCHRONIZATION
REFERENCES
Media is one of the important components for in vitro cultivation of animal cells. Every animal cells have specific requirements and media are designed by keeping in mind those requirements. However, the basic components and design principle remains the same. Every cell culture media contain carbon source, nitrogen source, trace elements, pH indicator, antibiotics ( although it is not recommended) for high value cell culture applications. While designing media various aspects are considered such as availability, cost effectiveness, types off cells to be grown and regulatory requirements. Tis slide also contains sample MCQs questions
Cell synchronization helps in obtaining distinct sub population of cells representing different stages of cell cycle.It helps in collecting population wide data of cells progressing through various stages of cell cycle. Immortalization, refers to cells having capability of undergoing cell division infinitely. Immortal cells are particularly preferred in cell culture to enable long time storage and use. This presentation teaches about cell synchronization, methods of cell synchronization, cellular transformation, immortalization and mechanism of immortalization.
For decades, cell lines have played a critical role in scientific developments. In most cases, researchers just got data generated from cell lines. However, due to some weaknesses of cell lines, scientists become increasingly cautious about these generated results. But now the game has changed! Primary cells now are believed to be a more biologically relevant tool than cell lines for studying human and animal biology. And we design this primary cell culture guide aimed at showing new investigators the basic principles of primary cell and some practical culture skills.
Primary and established cell line cultureKAUSHAL SAHU
Introduction
Primary Culture
Steps of Primary Culture
Isolation Of Tissue
Dissection And Disaggregation
Types Of Primary Culture
Primary Explants Culture
Enzymatic Disaggregation
Mechanical Disaggregation
Cell Line( Finite & Continuous)
Naming A Cell Line
Choosing A Cell Line
Maintenance Of Cell Line
Conclusion
Reference
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
SYNCHRONOUS CULTURES CAN BE OBTAINED IN SEVERAL WAYS:
Physical fractionation .
Chemical appro ach
CENTRIFUGAL ELUTRIATION
Inhibition of DNA synthesis
Nutritional deprivation
SYNCHRONIZATION AT LOW TEMPERATURE
CELLULAR TOTIPOTENCY
SOME HIGHLIGHTS OF CELL SYNCHRONIZATION
REFERENCES
Media is one of the important components for in vitro cultivation of animal cells. Every animal cells have specific requirements and media are designed by keeping in mind those requirements. However, the basic components and design principle remains the same. Every cell culture media contain carbon source, nitrogen source, trace elements, pH indicator, antibiotics ( although it is not recommended) for high value cell culture applications. While designing media various aspects are considered such as availability, cost effectiveness, types off cells to be grown and regulatory requirements. Tis slide also contains sample MCQs questions
Cell synchronization helps in obtaining distinct sub population of cells representing different stages of cell cycle.It helps in collecting population wide data of cells progressing through various stages of cell cycle. Immortalization, refers to cells having capability of undergoing cell division infinitely. Immortal cells are particularly preferred in cell culture to enable long time storage and use. This presentation teaches about cell synchronization, methods of cell synchronization, cellular transformation, immortalization and mechanism of immortalization.
For decades, cell lines have played a critical role in scientific developments. In most cases, researchers just got data generated from cell lines. However, due to some weaknesses of cell lines, scientists become increasingly cautious about these generated results. But now the game has changed! Primary cells now are believed to be a more biologically relevant tool than cell lines for studying human and animal biology. And we design this primary cell culture guide aimed at showing new investigators the basic principles of primary cell and some practical culture skills.
Applications of fish celllines by B.pptxB. BHASKAR
Recent research studies on fish cell lines found many more application of cell lines pathological studies, toxicology, biomedical research, vaccine development etc
Viruses are obligate intracellular parasites which means they can only grow or reproduce inside a host cell.
The primary purpose of virus cultivation:
To isolate and identify viruses in clinical samples.
To do research on the viral structure, replication, genetics, and effects on the host cell.
To prepare viruses for vaccine production.
Isolation of the virus is always considered a gold standard for establishing the viral origin of the disease
topics covered
CULTIVATION OF VIRUSES
Animal inoculation
Embryonated eggs
CAM
Allantoic cavity
Amniotic cavity
Yolk sac
Tissue culture
Organ culture
Explant culture
Cell culture
Primary cell culture
diploid cell culture
Continues cell lines
Compiled by Nagendra P and Pritam Vishu Bagwe
M.Tech Pharmaceutical Sciences
Department of Pharmaceutical Sciences and Technology
Institute of Chemical Technology, Matunga, Mumbai, India.
Vero cells are the continuous cell lines which is employed in the production of viral vaccines . This cell line has the ability to be scaled up and grown in large bioreactors using microcarrier beads .
Neuro-symbolic is not enough, we need neuro-*semantic*Frank van Harmelen
Neuro-symbolic (NeSy) AI is on the rise. However, simply machine learning on just any symbolic structure is not sufficient to really harvest the gains of NeSy. These will only be gained when the symbolic structures have an actual semantics. I give an operational definition of semantics as “predictable inference”.
All of this illustrated with link prediction over knowledge graphs, but the argument is general.
Elevating Tactical DDD Patterns Through Object CalisthenicsDorra BARTAGUIZ
After immersing yourself in the blue book and its red counterpart, attending DDD-focused conferences, and applying tactical patterns, you're left with a crucial question: How do I ensure my design is effective? Tactical patterns within Domain-Driven Design (DDD) serve as guiding principles for creating clear and manageable domain models. However, achieving success with these patterns requires additional guidance. Interestingly, we've observed that a set of constraints initially designed for training purposes remarkably aligns with effective pattern implementation, offering a more ‘mechanical’ approach. Let's explore together how Object Calisthenics can elevate the design of your tactical DDD patterns, offering concrete help for those venturing into DDD for the first time!
UiPath Test Automation using UiPath Test Suite series, part 3DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 3. In this session, we will cover desktop automation along with UI automation.
Topics covered:
UI automation Introduction,
UI automation Sample
Desktop automation flow
Pradeep Chinnala, Senior Consultant Automation Developer @WonderBotz and UiPath MVP
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
Dev Dives: Train smarter, not harder – active learning and UiPath LLMs for do...UiPathCommunity
💥 Speed, accuracy, and scaling – discover the superpowers of GenAI in action with UiPath Document Understanding and Communications Mining™:
See how to accelerate model training and optimize model performance with active learning
Learn about the latest enhancements to out-of-the-box document processing – with little to no training required
Get an exclusive demo of the new family of UiPath LLMs – GenAI models specialized for processing different types of documents and messages
This is a hands-on session specifically designed for automation developers and AI enthusiasts seeking to enhance their knowledge in leveraging the latest intelligent document processing capabilities offered by UiPath.
Speakers:
👨🏫 Andras Palfi, Senior Product Manager, UiPath
👩🏫 Lenka Dulovicova, Product Program Manager, UiPath
The Art of the Pitch: WordPress Relationships and SalesLaura Byrne
Clients don’t know what they don’t know. What web solutions are right for them? How does WordPress come into the picture? How do you make sure you understand scope and timeline? What do you do if sometime changes?
All these questions and more will be explored as we talk about matching clients’ needs with what your agency offers without pulling teeth or pulling your hair out. Practical tips, and strategies for successful relationship building that leads to closing the deal.
Essentials of Automations: Optimizing FME Workflows with ParametersSafe Software
Are you looking to streamline your workflows and boost your projects’ efficiency? Do you find yourself searching for ways to add flexibility and control over your FME workflows? If so, you’re in the right place.
Join us for an insightful dive into the world of FME parameters, a critical element in optimizing workflow efficiency. This webinar marks the beginning of our three-part “Essentials of Automation” series. This first webinar is designed to equip you with the knowledge and skills to utilize parameters effectively: enhancing the flexibility, maintainability, and user control of your FME projects.
Here’s what you’ll gain:
- Essentials of FME Parameters: Understand the pivotal role of parameters, including Reader/Writer, Transformer, User, and FME Flow categories. Discover how they are the key to unlocking automation and optimization within your workflows.
- Practical Applications in FME Form: Delve into key user parameter types including choice, connections, and file URLs. Allow users to control how a workflow runs, making your workflows more reusable. Learn to import values and deliver the best user experience for your workflows while enhancing accuracy.
- Optimization Strategies in FME Flow: Explore the creation and strategic deployment of parameters in FME Flow, including the use of deployment and geometry parameters, to maximize workflow efficiency.
- Pro Tips for Success: Gain insights on parameterizing connections and leveraging new features like Conditional Visibility for clarity and simplicity.
We’ll wrap up with a glimpse into future webinars, followed by a Q&A session to address your specific questions surrounding this topic.
Don’t miss this opportunity to elevate your FME expertise and drive your projects to new heights of efficiency.
Builder.ai Founder Sachin Dev Duggal's Strategic Approach to Create an Innova...Ramesh Iyer
In today's fast-changing business world, Companies that adapt and embrace new ideas often need help to keep up with the competition. However, fostering a culture of innovation takes much work. It takes vision, leadership and willingness to take risks in the right proportion. Sachin Dev Duggal, co-founder of Builder.ai, has perfected the art of this balance, creating a company culture where creativity and growth are nurtured at each stage.
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
Connector Corner: Automate dynamic content and events by pushing a buttonDianaGray10
Here is something new! In our next Connector Corner webinar, we will demonstrate how you can use a single workflow to:
Create a campaign using Mailchimp with merge tags/fields
Send an interactive Slack channel message (using buttons)
Have the message received by managers and peers along with a test email for review
But there’s more:
In a second workflow supporting the same use case, you’ll see:
Your campaign sent to target colleagues for approval
If the “Approve” button is clicked, a Jira/Zendesk ticket is created for the marketing design team
But—if the “Reject” button is pushed, colleagues will be alerted via Slack message
Join us to learn more about this new, human-in-the-loop capability, brought to you by Integration Service connectors.
And...
Speakers:
Akshay Agnihotri, Product Manager
Charlie Greenberg, Host
Key Trends Shaping the Future of Infrastructure.pdfCheryl Hung
Keynote at DIGIT West Expo, Glasgow on 29 May 2024.
Cheryl Hung, ochery.com
Sr Director, Infrastructure Ecosystem, Arm.
The key trends across hardware, cloud and open-source; exploring how these areas are likely to mature and develop over the short and long-term, and then considering how organisations can position themselves to adapt and thrive.
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GraphRAG is All You need? LLM & Knowledge GraphGuy Korland
Guy Korland, CEO and Co-founder of FalkorDB, will review two articles on the integration of language models with knowledge graphs.
1. Unifying Large Language Models and Knowledge Graphs: A Roadmap.
https://arxiv.org/abs/2306.08302
2. Microsoft Research's GraphRAG paper and a review paper on various uses of knowledge graphs:
https://www.microsoft.com/en-us/research/blog/graphrag-unlocking-llm-discovery-on-narrative-private-data/
3. History
• History
• The 19th-century English physiologist Sydney Ringer developed salt solutions
containing the chlorides of sodium, potassium, calcium and magnesium
suitable for maintaining the beating of an isolated animal heart outside of the
body.[1] In 1885, Wilhelm Roux removed a portion of the medullary plate of
an embryonic chicken and maintained it in a warm saline solution for several
days, establishing the principle of tissue culture.[3]
Ross Granville Harrison,
working at Johns Hopkins Medical School and then at Yale University,
published results of his experiments from 1907 to 1910, establishing the
methodology of tissue culture.[4]
• Cell culture techniques were advanced significantly in the 1940s and 1950s to
support research in virology. Growing viruses in cell cultures allowed
preparation of purified viruses for the manufacture of vaccines. The injectable
polio vaccine developed by Jonas Salk was one of the first products mass-
produced using cell culture techniques. This vaccine was made possible by the
cell culture research of John Franklin Enders, Thomas Huckle Weller, and
Frederick Chapman Robbins, who were awarded a Nobel Prize for their
discovery of a method of growing the virus in monkey kidney cell cultures.
3Note:-The neural plate is a key developmental structure that serves as the basis for the nervous
system
4. Introduction:
• Cell culture is the complex process by which cells are grown under
controlled conditions, generally outside of their natural environment.
In practice, the term "cell culture" now refers to the culturing of cells
derived from multi-cellular eukaryotes, especially animal cells.
However, there are also cultures ofplants, fungi and microbes,
including viruses, bacteria and protists. The historical development
and methods of cell culture are closely interrelated to those of
tissue culture and organ culture.
4
5. 5
Purpose
Cell culture is essential for growth of viruses because they
are obligate intracellular parasites; they cannot replicate in
any cell-free medium, and they require living cells from a
suitable host within which to multiply. Animals such as mice
and embryonated avian eggs may be used for the propagation
of viruses, but for various reasons (time, cost, uniformity,
ease of handling, animal welfare, etc.), the propagation of
most viruses in cultured living cells is the method of choice
today.
Cell culture is also used in recombinant DNA technology
and many in vitro assays.
6. Isolation
• Cells can be isolated from tissues for ex vivo culture in several ways.
Cells can be easily purified from blood; however, only the white cells
are capable of growth in culture. Mononuclear cells can be released
from soft tissues by enzymatic digestion with enzymes such as
collagenase, trypsin, or pronase, which break down the
extracellular matrix. Alternatively, pieces of tissue can be placed in
growth media, and the cells that grow out are available for culture.
This method is known as explant culture.
Collagenase Collagenases are enzymes that break the peptide bonds in collagen.
Pronase is a commercially available mixture of proteinases isolated from the extracellular fluid of Streptomyces griseus.
Activity extends to both denatured and native proteins leading to complete or nearly complete digestion into
individual amino acids.
tripsin is a serine protease found in the digestive system of many vertebrates, where it hydrolyses proteins.
6
8. 8
Primary cell cultures.
When cells are taken freshly from animal tissue and placed in
culture, the cultures consist of a wide variety of cell types, most
of which are capable of very limited growth in vitro, usually
fewer than ten divisions. These cells retain their diploid
karyotype, i.e., they have the chromosome number and
morphology of their tissues of origin. They also retain some of
the differentiated characteristics that they possessed in
vivo. Because of this, these cells support the replication of a
wide range of viruses. Primary cultures derived from monkey
kidneys, mouse fetuses, and chick embryos are commonly used
for diagnostic purposes and laboratory experiments.
a cell line derived directly from the parent tissue.
Cells in primary culture have the same karyotype
and chromosome number as those in the original
tissue.
Can be subculture only one or twice e.g-primary
monkey, baboon kidney
9. 9
2. Diploid cell strains.
Which are derived from human fetal tissue and can be
subcultured 20-50 times e.g-human diploid fibroblast such as
MRC-5(human fetal lung fibroblast establish from normal lung
tissue of 14 week old male foetus)
Some primary cells can be passed through secondary and
several subsequent subcultures while retaining their original
morphological characteristics and karyotype. Subcultures will
have fewer cell types than primary cultures. After 20 to 50
passages in vitro, these diploid cell strains usually undergo a
crisis in which their growth rate slows and they eventually die
out. Diploid strains of fibroblasts derived from human fetal
tissue are widely used in diagnostic virology and vaccine
production.
10. 3. Continuous cell lines. Derived from tumors of human or animals
e.g- Vero, Hep2
Certain cultured cells, notably mouse fetal fibroblasts, kidney cells from various
mammalian species, and human carcinoma cells, are able to survive the growth crisis
and undergo indefinite propagation in vitro. After several passages, the growth rate
of the culture slows down; then isolated colonies of cells begin to grow more rapidly
than diploid cells, their karyotype becomes abnormal (aneuploid), their morphology
changes, and other poorly understood changes take place that make the cells
immortal. The cells are now "dedifferentiated," having lost the specialized
morphology and biochemical abilities they possessed as differentiated cells in vivo.
Continuous cell lines such as KB and HeLa, both derived from human carcinomas,
support the growth of a number of viruses. These lines and others derived from
monkey kidneys (e.g., Vero), mouse fetuses (L929), and hamster kidneys (BHK) are
widely used in diagnostic and experimental virology. Continuous cell lines have been
established from many types of vertebrate and invertebrate animal tissues and are
available from the American Type Culture Collection.
11. 11
Primary Cell Culture
• Sterilization of
Cubicle
• Preparation of Media
• Preparation of
dispersion solution
• Preparation of
Monolayers
12. 12
Sterilisation of Cubicle (compartment)
• Fumigation
• 35 ml of formaline and 17.5 g of potassium
permanganate per 100 cubic feet of air space
• Keep the cubicle closed for 65 hours
• Working Surface Sterilization
• Mop with 5% phenyl/ 1/40 solution of dettol
• Use 1 % Copper sulphate for fungal disinfection
13. 13
Cell Culture Media
• Balanced Salt Solution (BSS)
• Glucose
• Amino acids
• Vitamins
• Sodium bicarbonate
• Phenol red
• Fetal Calf Serum
14. 14
Culture Media
Prepare stock antibiotic solution
Weigh the required amount of Ready made Balanced Salt Medium and
add sterile water
Add the required amount of antibiotic stock solution into BSS
Adjust the pH of the solution using 2.8% / 7.5% Sodium bicarbonate
or 0.1 N Hydrochloric acid to 7.4
Sterilize using 0.2 µ Seitz filter applying positive pressure.
Add sterile heat inactivated fetal calf serum to make a final
concentration of 15%
Store at 4o
C.
15. 15
Dispersion solution
• Weigh Calcium Magnesium free Buffer premix and add sterile water
• Add 0.1 % w/v Trypsin powder
• Adjust pH to 7.4 to 7.6
• Sterilize by Filtration through Seitz filter
• Store at - 20o
C
17. 17
Materials
Ten to Eleven days Embryonated Chicken Eggs
Forceps, Scissors, Rubber bulbs
Pipettes, Beakers, Centrifuge tubes, Flasks, Petri dishes, Trypsinisation flask
Seitz filter assembly, Vacuum/air pump
Mono-pan analytical balance
pH meter
Magnetic stirrer, Teflon coated magnetic stirrer, Cyclo-mixer
Refrigerated Centrifuge
Laminar Air Flow Apparatus
18. 18
Preparation of Chicken Embryo Fibroblast MonolayerPreparation of Chicken Embryo Fibroblast Monolayer
1. Sterilize Outer shell of the Eggs
2. Remove the Shell, Shell membrane and CAM
3. Wash embryo with BSS
4. Remove Head, Limbs and Internal Organs
5. Wash several times with BSS
6. Cut into small pieces of ~ 1mm thickness
7. Wash in BSS
8. Trypsinize /.
9. Filter with a cheese cloth
10. Centrifuge and pack cells
11. Resuspend in growth medium
12. Repeat step 10 and 11, two more times
13. Count the cells and adjust to a concentration of 10 6
cells per ml
14. Seed Culture tubes/ flasks/ Bottles and Incubate at 37 o
C for 48 to 72
hours.
19. 19
procedure
• Disinfect the surface of the egg
over the air sac. With scissors
or the blunt-end of a forceps,
break the shell over the air sac
• Opening of egg through a
circular incision around the
airsac
•10- to 12-day-old embryonated eggs
34. 34
• Preservation of Cultured Cells by Freezing
• 50 to 80% of the cells of a healthy culture will
survive freezing.
• . Cells must be stored at -70o
C or lower
35.
36. 36
Ex vivo (Latin: out of the living) means that which takes place outside an organism. In science, ex vivo refers
to experimentation or measurements done in or on living tissue in an artificial environment outside the
organism with the minimum alteration of the natural conditions. The most common "ex vivo" procedures
involve living cells or tissues taken from an organism and cultured in a laboratory apparatus, usually under
sterile conditions and no alterations done for a few hours up to 24 hrs. Experiments lasting longer than this
using living cells or tissue are typically considered to be "in vitro". Ex-vivo conditions allow experimentation
under highly controlled conditions impossible in the intact organism, albeit at the expense of looking at the
tissue in its "natural" environment. One widely performed ex vivo study is the chick chorioallantoic
membrane (CAM) assay. In this assay, angiogenesis is promoted on the CAM membrane of a chick embryo
outside the organism (chicken). Ex vivo studies are usually performed in vitro, although the use of these two
terms is not synonymous.
In vitro (Latin for within the glass) refers to the technique of performing a given procedure in a controlled
environment outside of a living organism. Some may argue that in vitro refers to a process that is created in a
"test tube"; however, Robert Kail and John Cavanaugh on page 58 in the 4th edition of Human Development:
A Life-Span View cite that in fact the process is contained in a petridish. Many experiments in cellular
biology are conducted outside of organisms or cells; because the test conditions may not correspond to the
conditions inside of the organism, this may lead to results that do not correspond to the situation that arises in
a living organism. Consequently, such experimental results are often annotated with in vitro, in
contradistinction with in vivo.
What's the difference between ex vivo and in vitro?
Or is it the same thing
37. Institute of Animal Health and
Veterinary Biologicals
37
karyotype
the chromosomal constitution of the cell nucleus; by extension, the photomicrograph of
chromosomes arranged
38. 38
Diploid cultures vs. continuous cell lines
What is the difference between a diploid culture and a
continuous cell line?
-Diploid cultures have a finite lifespan. They can undergo a
maximum of 20-80 PDLs (Population Doubling Level) before
they senesce. Normal human cells, such as human skin
fibroblasts, are one example of diploid cells.
-Continuous cell lines are immortalized cell lines with an infinite
lifespan. These usually either come from tumor tissue or have
been deliberately immortalized or transformed. However, many
rodent cell lines spontaneously transform.
39. 39
Formulations of Media and Solutions
Saline A
Ingredient g/l
NaCl 8.0
KCl 0.4
NaHCO3 0.35
Glucose 1.0
Phenol red 0.05
Add distilled H2O to 1 liter. Filter sterilize. Saline A is usually prepared as a
10X solution and stored at -20o
C.
Saline-Trypsin-EDTA (STE)
Ingredient g/l
Trypsin (1:250) 0.5
EDTA (disodium salt) 0.2
NaCl 8.0
KCl 0.4
NaHCO3 0.35
Glucose 1.0
Phenol red 0.05
Add distilled H2O to 1 liter. Dissolve trypsin first by slowly stirring in ~200
ml H2O (avoid denaturation). Add remaining ingredients and filter sterilize.