vitrification in ivf
DESCRIPTION
Clinical overview of freezing of embryo using vitrificationTRANSCRIPT
VITRIFICATION
Dr G A RAMA RAJU
Over viewComponents of
cryobiology
Slow freezing /vitrification
Key issues in
vitrification
sperm
EmbryoOocyt
e
Ovary /testis
Safety
New born
.
Prof Hans Tinneberg
Pioneers.
Nature
?
? A vitrified frog felt as a rock
A wood frog in winter - Alaska. Nature’s expt with VITRIFICATION
Cell injury.
1 Cell size shape ,permeability ,individual quality and sensitivity.2 cryoprotectant toxicity3 cooling and warming.
The land mark publication “Slow cooling” Trounson and Mohr 1983
A long gap…………………………………….
PLANER COMPUTERISED FREEZER
Vitrification a tipping point in ART
Gopubmed:
Father Basile.j.luyet
work between 1937-1958
1952 Time magazine
Innovator :
Spermatozoa : Early adopter
15
Early adopter: PARKES
Majority
Publication from different countries
CRYOPRESRVATIOON PROCESSES
•Add cryoprotectant
First Concept
•Cool cells to -196 & store
Second Concept
•Warm/Thaw
Third Concept
•Remove cryoprotectant
Fourth concept.
Disadvantage and benefits of vitrification
ResultsTimeCost
vitrification
Slow freezing
*
Variables Variable that effect vitrificationof vitrification that can profoundly influence its effectiveness
19
Summary of the primary benefits of vitrification
20Biology of Reproduction December 1, 2002 vol. 67 no. 6 1671-1680
Over view of current work.
DMSOPROPYLENE GLYCOLGLYCEROL
OTHER SACCHARIDESOSMOTIC ALTERATION
CPA
HEPES BUFFERED MEDIA
VITRIFICATION SOLUTIONS
Are all vitrification and warming solution same ?
Yes and No
MOPS
Sol-A: 7.5% of EG +
7.5% of DMSO
Sol-B: 15% of EG + 15% of
DMSO + 1M of SUCROSE + FICOLL (10 mg/ml)
ES: 7.5%
of EG
+ 7.5% of
DMSO
VS: 15% of EG + 15% of DMSO +
0.6M of SUCROSE
Sage Bio Pharma
V1:
BUFFER
V2: EG + Propanediol
V3: EG + Propanediol + FICOLL
Vitrolife
ES: 7.5%
of EG
+ 7.5% of 1,2
Propanediol
VS: 15% of EG + 15% of 1,2
Propanediol + 0.5M of SUCROSE
Medi Cult
Rama Raju et al
Composition of Vitrification Solutions
1M Sucrose
0.5M Sucrose
0.25M Sucrose
1M Sucrose
0.5M Sucrose
Sage Bio Pharma
W1:
Sucrose
W2: Sucrose
W3: Sucrose
Vitrolife
Medi Cult
0.125M Sucrose
1M Sucrose
0.5M Sucrose
0.25M Sucrose
Rama Raju et al
Composition of Warming Solutions
Carrier
Cryoloop,cryotip,pulled straw, electronic grid
Vitrification and slow freezing similar results
Japanese group
vajata
We are working on removal of water from
oocyte and working on that
Vitrification of 2 pronuclei
DAY 2 OR DAY 3 EMBRYO
33
Copyright restrictions may apply.
Devroey, P. et al. Hum Reprod Update 2009 15:391-408; doi:10.1093/humupd/dmp012
The odds of post-thawing survival rate of cleavage-stage embryos after vitrification or slow freezing
Our first vitrification twins
Blastocyst
38
Blastocyst expanded or hatched
39
Safety issues
Ultraviolet light to sterilize the liquid nitrogen
IVF children
Use of hospital services
Long hospitalizatio
n
Increase risk of infection epilepsy
Cerebral palsy
Sleep disturbances Convulsions Behavior
problems
Congenital malformation
Need for long tern follow up to look deeply
Safe?
cryopreservation children
Need for long tern follow up to look deeply
?
Perinatal outcomes in vitrified oocytes
Neonatal outcome of day 3 embryos
Perinatal outcome of vitrificaion blastocyst
acknowledgement Hans Tinneberg my mentor
summaryVisakhapatnam Beach .