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.

1. CRYOPRESERVATION
SPERM BANKING
AND
ARTIFICIAL INSEMINATION
G. Karthikeyan
LS14108
2nd Year
M.Sc., Life Sciences
Bharathidasan University

2. INTRODUCTION
 Cryobiology derives from three greek words:
 Cryos- cold; Bios-Life; logos-course of study
 It is defined as the process of arresting all
biological process and placing biological
material (cells, oocytes, embryos,
spermatozoa or tissue )into a suspended
state of animation at an ultra-low
temperature.

4. CRYOPRESERVATION
 First documented by Boyle (1663).
 First concept of sperm preservation by
Lazzaro spallanzini(1760).
 Most significant discovery was the use of
glycerol as cryoprotectant by Polge and
smith(1949)

5. PRINCIPLE
1. FREEZING
2. STORING
3. THAWING

6. CONT..
 The ability of glycerol to protect cells from freezing
injury was discovered accidentally.
 The subsequent development of cryopreservation
techniques has had a huge impact in many fields,
most notably in reproductive medicine.


7.  Freezing injury has been shown to have two
components, direct damage from the ice crystals
and secondary damage caused by the increase in
concentration of solutes as progressively more ice is
formed.
 Intracellular freezing is generally lethal but can be
avoided by sufficiently slow cooling, and under
usual conditions solute damage dominates.

8. WHY CRYOPRESERVATION?
 Availability and preservation of raw biological
materials are the major challenge in the
widespread use of assisted reproduction
technology in humans and livestock.
 For example, many artificial insemination
procedures rely on availability of cryopreserved
sperm.
 Similarly, cryopreserved oocytes and embryos are

9. CRYOPROTECTANTS
 A cryoprotective agent is a substance added to the
freezing medium to prevent the biological material
from freezing injures that is damage due to ice
formation
 In experimental biology, the conventional CPAs are
glycols and glycerol
Common cryoprotectants:
 DMSO-Dimethyl sulfoxide
 Ethylene glycol
 Glycerol

10.  The presence of cryoprotectants in the freezing solution is
necessary to prevent cell damage during freezing and
thawing of the samples
 Cryoprotectants are included in the solutions to freeze
the cells to lower the freezing point
CONT..

11. SPERM BANKING
 Sperm banking is the name for the
collection and storage of semen
 Semen is the fluid that contains sperm,
Sperm banking is also known as sperm
cryopreservation or semen storage

13. PURPOSE:
 The purpose of cryopreserving semen
(sperm banking) is to help ensure the
possibility of conception in the future
TWO TYPES :
 Short-term Semen Cryobanking
 Long-Term Semen Cryobanking

14. Short-term Semen Cryobanking
 Short-term semen cryobanking is the depositing, freezing
and storage of sperm at a sperm bank for less than one year
 Cryobanked sperm is then used in artificial insemination, in
vitro fertilization (IVF) and other fertility treatment
procedures
 Assisted reproductive technologies ,
(i.e., in vitro fertilization, gamete intrafallopian transfer, etc...)

15. Long-Term Semen
Cryopreservation
 Time-tested techniques enable semen
specimens and embryos to be frozen and
stored indefinitely in liquid nitrogen
 According to the donor’s specific wishes,
these specimens and embryos can later be
thawed and used in an attempt to
conceive through artificial insemination,
IVF or other fertility treatment techniques.

16. ARTIFICIAL INSEMINATION
 Artificial insemination is a fertility treatment
that delivers sperm to the female using a tube
to reach the cervix for intracervical
insemination (ICI) or the uterus for intrauterine
insemination (IUI)
 ICI and IUI are very quick and painless
procedures that can be attempted many
times without any injury to the receiving

18. Artificial Insemination Method
 The insemination procedure involves
injecting prepared sperm into the female
reproductive tract through the vagina
 Intra cervical insemination (depositing of
sperm into the cervical opening)
 Intra uterine insemination (depositing of
sperm into the uterus)

20.  Speculum , a medical tool is used to insert the sperms
into female reproductive track
 Small amount of fluid is present inside the injection
needle were the catheter is at edge of the needle
Needle is fixed into speculum and further process of
injection was carried out
 The insemination procedure takes 5 to 10 minutes and
is usually performed once or twice a month until
pregnancy is achieved
Procedure

22. REFERENCE
• Textbook of Animal Biotechnology
• Institute of Cancer Biology , United
Kingdom

23. Tq