Vitrification of oocytes ppt

Solidification process in which oocytes are treated with cryo-protective substance and submerged in liquid nitrogen at a temperature of -1960C

1. Tracking programme
On
Vitrification of oocytes

2. Introduction
Solidification process in which oocytes are treated
with cryo-protective substance and submerged in
liquid nitrogen at a temperature of -196
0
C.
It is the method done for the cryopreservation of
oocytes.
Rapid/fast freezing or non-equilibrium type
method .
Firstly used by the Greg Fahy and William, F.Rall
in 1985.

3. Advantage of oocyte collection
Use of superior quality of female
Avoid problems of ageing in older females
Oocyte donation
Options for delayed motherhood
Avoid ethical issue and legal restrictions
related to embryo banking

4. Methods of collection of oocytes
1. Aspiration method
2. Slicing
3. Dissecting

5. Aspiration method
Most efficient technique to obtain
oocytes.
OPU (Ova pick up) by puncturing of
ovarian follicles by leproscopy ultra-
sound.
OPU provides 4-8 oocytes per collection
should be done weekly interval from
adults.

6. Preservation of oocytes
Done with the help of cryo-protectants namely:
 Glycerol,
 DMSO,
 Liquid nitrogen (-196
0
C)
 Propanediol

7. Cryotech vitrification kit
Material :
• Warming solution(TS):1vial of 1.8 ml
• Diluents solution(DS):1vial of 0.5 ml
• Washing solution(WS):1vial of 1 ml
• 1 warm plate with 4 wells
• Microscope
• Stop watch
• Tweezers
• Micro pipette

8. 1) Preparation:
Place the warm plate and TS vial in the incubator at 37
0
C > 3 hr.
Bring DS and WS vials to room temperature at least 1 hr.
Prepare fresh liquid nitrogen.

9. 2) Warming (1 min):
•Quickly with in 1 sec put the cryotech into TS
well
•Start stop watch for 1 min
•Oocyte separate from the cryotech sheet by itself
and begin to floats.

10. 3) Dilution (3 min):
Aspirate the oocyte 3 mm long DS in to the pipette.
Blow out DS into the bottom center of DS and expel the
oocyte slowly bottom of the TS layer in DS well.
4) Washing 1 (5 min)
5) Washing 2 (1 min)

11. Conclusion
•Oocyte could not be successfully frozen for
more than 20 yrs because its high
cryosensitivity.
•But today any one can preserve it easily and
with 99% of survival by cryotech method.

12. Thank you