An ideal oocyte activation protocol and embryo culture conditions for somatic cell nuclear transfer using sheep oocytes.

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dc.contributor.authorPatel, Hiren
dc.contributor.authorChougule, Shruti
dc.contributor.authorChohan, Parul
dc.contributor.authorShah, Naval
dc.contributor.authorBhartiya, Deepa
dc.date.accessioned2014-12-12T08:38:58Z
dc.date.available2014-12-12T08:38:58Z
dc.date.issued2014-10
dc.description.abstractPluripotent stem cells are possibly the best candidates for regenerative medicine, and somatic cell nuclear transfer (SCNT) is one of the viable options to make patient-specific embryonic stem cells. Till date efficacy of SCNT embryos is very low and requires further improvement like ideal oocyte activation and in vitro culture system. The aim of the present study was to evaluate ideal oocyte activation using different stimulation protocols and to study the effect of cumulus co-culture conditions on embryo development. Results demonstrate that between electric stimulation and chemical stimulation using calcium ionomycin and ionophore, best oocyte activation was obtained using calcium ionomycin (5 μM for 5 min) which resulted in 83% cleavage followed by 7% of early blastocyst which further increased to 15% when a cumulus bed was also introduced during embryo culture. Sequential modified Charles Rosenkrans 2 (mCR2) medium was used for embryo culture in which glucose levels were increased from 1 mM to 5 mM from Day 3 onwards. SCNT using cumulus cells as donor somatic cell, calcium ionomycin to activate the reconstructed oocyte and embryo culture on a cumulus bed in sequential mCR2 medium, resulted in the development of 6% embryos to early blastocyst stage. Such technological advances will make SCNT a viable option to make patient-specific pluripotent stem cell lines in near future.en_US
dc.identifier.citationPatel Hiren, Chougule Shruti, Chohan Parul, Shah Naval, Bhartiya Deepa. An ideal oocyte activation protocol and embryo culture conditions for somatic cell nuclear transfer using sheep oocytes. Indian Journal of Experimental Biology. 2014 Oct; 52(10): 925-934.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/153779
dc.language.isoenen_US
dc.source.urihttps://nopr.niscair.res.in/handle/123456789/29436en_US
dc.subjectEmbryo cultureen_US
dc.subjectOocytesen_US
dc.subjectParthenogenesisen_US
dc.subjectSheepen_US
dc.subjectSomatic cell nuclear transferen_US
dc.subject.meshAnimals
dc.subject.meshBlastocyst --physiology
dc.subject.meshCell Culture Techniques --methods
dc.subject.meshEmbryonic Development --physiology
dc.subject.meshFemale
dc.subject.meshMale
dc.subject.meshNuclear Transfer Techniques
dc.subject.meshOocytes --physiology
dc.subject.meshParthenogenesis
dc.subject.meshSheep
dc.titleAn ideal oocyte activation protocol and embryo culture conditions for somatic cell nuclear transfer using sheep oocytes.en_US
dc.typeArticleen_US
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