Effect of in vitro culture media and assisted hatching techniques on mice embryo survival rate following cryopreservation

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dc.contributor.advisor Nedambale, T. L.
dc.contributor.advisor Fushai, F.
dc.contributor.author Serota, Nthabiseng Ruth
dc.date 2018
dc.date.accessioned 2018-06-06T09:24:10Z
dc.date.available 2018-06-06T09:24:10Z
dc.date.issued 2018-05-18
dc.identifier.uri http://hdl.handle.net/11602/1150
dc.description MSCAGR (Animal Science)
dc.description Department of Animal Science
dc.description.abstract This study determined the effects of in vitro culture media (Ham’s F10 and TCM199) and assisted hatching techniques (laser or mechanical) on mice embryo survival following cryopreservation. Pure strain C57BL/6 (B6) female (50) and strain BALB /c (C) Male (25) mice were crossed to produce F1 generation of females which were injected for follicular growth and super ovulation at 6 weeks of age and from which embryos were produced 21 h later through in vivo fertilization. Embryos were randomly divided into Petri dishes with different culture media, and the development of embryos was assessed until the morula stage. At the morula stage, selected embryos were assisted to hatch using different techniques, and then cryopreserved in liquid nitrogen using the slow freezing method for a period of 1 week. After 1 week of cryopreservation, the embryos were thawed and cultured in the two different in vitro culture media for 72 hours. Thereafter, the numbers of embryos hatched or survived were recorded after 24 h, 48 h and 72 h. Data was analyzed using ANOVA in Minitab Software Version 16 (2010). Significant difference in embryo quality development was observed between in vitro culture media and stage of embryo development (P<0.05). In the TCM-199 in vitro culture medium, embryo quality development yielded 72, 69 and 69% from day 1 to day 3, while in Ham’s F10 embryo quality development yielded 68, 63 and 60% respectively. Relative to the control (18.1%) assisted hatching improved hatchability significantly (P<0.05) in the order laser (23.6%)>, mechanical (20.8%). There was significant (P<0.01) interaction between assisted hatching techniques and evaluation time, whereby laser assisted hatching was most successful at 48 h (42.0%) while mechanical assisted hatching was most successful at 72 h (36.8%). Cryopreservation reduced the embryo survival compared to fresh embryos. In conclusion laser was the best assisted hatching technique, while TCM-199 was the better medium for in vitro culture of embryo. en_US
dc.description.sponsorship NRF en_US
dc.format.extent 1 online resource (x, 53 leaves : color illustrations)
dc.language.iso en en_US
dc.rights University of Venda
dc.subject Cryopreservation en_US
dc.subject Assisted hatching en_US
dc.subject Culture media en_US
dc.subject Embryo en_US
dc.subject.ddc 636.08245
dc.subject.lcsh Artificial insemination
dc.subject.lcsh Animal breeding
dc.subject.lcsh Reproductive technology
dc.subject.lcsh Mice as laboratory animals
dc.subject.lcsh Rodents
dc.title Effect of in vitro culture media and assisted hatching techniques on mice embryo survival rate following cryopreservation en_US
dc.type Dissertation en_US

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