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Cryopreservation Techniques and Applications
- 1. CRYOCRYO -PRESENTED BY -- DEEPSHIKHA KEOT ROLL NO. - 3 MSC. 1 ST SEM, DEPT. OF ZOOLOGY, GAUHATI UNIVERSITY
- 2. LET'S HAVE A GLIMPSE ON THE CONTENTLET'S HAVE A GLIMPSE ON THE CONTENT Sperm cryopreservation Embryo cryopreservation Cryonics ● INTRODUCTION ● MECHANISM / PROCEDURE OF CRYOPRESERVATION ● IMPORTANCE AND ITS APPLICATION ● REFERENCES ACKNOWLEDGEMENT
- 3. INTRODUCTIONINTRODUCTION CRYOPRESERVATION- ● Greek word ”krayos”--”frost”.Literally preservation in “frozen state”. ● Non- lethal, Freeze vital cells and tissues at very low (sub- zero) temperatures. RELATED CONCEPTS-RELATED CONCEPTS- ● Cryobiology—Invented in1964,literally-- ''cryo''=''cold'' and ''logos''=''science''. ● Cryogenics-- Branch of physics concerned with the study of causes and effects of extremely low temperatures. ● Cryonics---Branch of cryopreservation of clinically dead human body or an organ with a hope to restore it later with the help of future medical technologies.
- 4. HISTORICAL BACKGROUND MILESTONES OF SUCCESS HISTORICAL BACKGROUND MILESTONES OF SUCCESS Sl. no. YEAR OF DISCOVERY SCIENTIST DISCOVERY / WORK 1 1949 Ernest John Christopher Polge 1st person---solve the mystery-how to preserve cells and tissues at very low temperature 2 1950 James E. Lovelock Suggested –increasing salt concs. in a cell to lose water to the external ice might cause damage to the cell 3 1953 Jerome K.Sherman Successful freeze and thaw human sperm. Founded World's 1st Sperm Bank 4 1983 Alan Trounson Success achieved in pregnanancy after freezing early human embryos 1-3 days after fertilization 5 1986 Christopher Chen 1st----freeze and thaw human oocyte 6 1988 Yves Menezo 1st commercial culture media in invitro fertilisation. Blastocyst freeze and thaw 7 1993 Gianpiero Palermo 1st embryos after intra ICSI 8 1995 Edoward Servy and Zisu Liu 1st—transplant a cryopreserved blastocyst followed ICSI
- 5. NATURAL ACCESS IN CRYOPRESERVATIONNATURAL ACCESS IN CRYOPRESERVATION ● Cryopreservation in nature- Water bear Water bear Called the Tardigrade,are Microscopic and multicelullar 8 leged ,segmented micro- animals. Survive freezing by replacing most of internal water with sugar trehalose. Temperature they survive- ● A few min .at 151ºC ● 30 years at -20 ● A few days at -200 ● A few mins. At -272 ºC ºC ºC Water bear Arctic Salamenders Antarctic insects
- 6. MECHANISM OF CRYOPRESERVATIONMECHANISM OF CRYOPRESERVATION Steps involved--- 1. Selection and collection of the material1. Selection and collection of the material 3. Selection of cryoprotectant3. Selection of cryoprotectant 4.Freezing4.Freezing 2.Preparation of Extender solution2.Preparation of Extender solution 6. Thawing6. Thawing 5.Storage5.Storage 8. Determination of Survival/Viability8. Determination of Survival/Viability 7. Washing and reculture7. Washing and reculture No. of cells growing/no. of cells thawed*100 Mounibs medium(M) Menezo medium(Me) --Hank's BSS---- In addition ,nutrients ,glycine, antibiotics. +40ºC 40-50 0 -79 -139 -196 Dehydration freezing Penetrating Non-Penetrating ºC ºC ºC ºC ºC Slow freezing Step-wise freezing Rapid freezing Diluent ● Advantages ● Disadvantages Prevention of risks Ice crystal formation Dehydration Shocks Slow freezing Vitrification
- 7. IMPORTANCE AND APPLICATIONIMPORTANCE AND APPLICATION cell organ tisssuegametes individuals Applicable in- ● Research work ● Biodiversity conservation ● Medicine ● Biotechnology ● Forestry ● Wildlife ● Plant germplasm ● Genetics and fertilisation ● genetic resourses ● endangered species preservation ● conventional method of storage failed ● long term preservation of material.
- 8. CRYONICS (HUMAN BODY PRESERVATION) CRYONICS (HUMAN BODY PRESERVATION) ● Cryonics—Greek'' kayos- cold'', is the low temperature preservation(@ 196 C) of people who cannot be sustained by contemporary medicine, with the hope that resuscitation and restoration to full health may be possible in the far future. ● A 14 yr old girl with terminal cancer won the right to have her body cryopreserved.
- 9. CRYONICS (HUMAN BODY PRESERVATION) CRYONICS (HUMAN BODY PRESERVATION) ● Cryonics—Greek'' kayos- cold'', is the low temperature preservation(@ 196 C) of people who cannot be sustained by contemporary medicine, with the hope that resuscitation and restoration to full health may be possible in the far future. ● A 14 yr old girl with terminal cancer won the right to have her body cryopreserved.
- 10. REFERENCESREFERENCES ● Dr.R.C Dubey ,''A textbook of Biotechnology''published by S.Chand. ● B.D.Singh( 2004) ''Biotechnology Expanding Horizons'' published by Kalyani Publishers,New Delhi-110002. ● Www.fao.org>docrep ● https://www.biw.kuleuven.be>tro>COST ● https://www.slideshare.net ● https://linkdin.com ● https://m.wikihow.com
- 11. ACKNOWLEDGEMENTACKNOWLEDGEMENT At the very outset,I would like to extend my sincere gratitude to Prof.Jatin Kalita sir (HOD),Dept. Of Zoology,G.U. for giving us such an opportunity to talk before a litterate crowd and test ourselves in presenting a seminar. I even thank wholeheartly to Prof.Jogen Chandra Kalita sir,Dept. Of Zoology,G.U for his inspiring and motivated speech who guided very efficient in my presntation.I thank and acknowledge my deep sense of gratitude to Prof. Manas Das sir,Dept. Of Zoology,G.U for organising the seminar and guiding us with our topics. I even thank my well-wishers for their encouragement.Lastly, I thank my parents for allowing me to pursue higher education and encouraging me in all my way. Yours Obediently Deepshikha Keot, Roll No. -3 M.Sc. 1St sem, Dept. of Zoology, Gauhati University.