NDRI Quinquennium Book COVER FINAL CURVE.cdr

67 results in the cow producing beta casein A1 while theproline results in the cow producing beta casein A2.Biotechnological Reproductive and MolecularTechnologies...••Interspecies blastocyst stage embryos were producedby Handguided cloning using buffalo cytoplasts anddifferentiated somatic cells from cattle and goat.••Zona-free buffalo oocytes were successfully activated forparthenogenetic development for producing blastocyststage embryos using chemical or electrical stimulation.••Cloning efficiency was shown to be improved by increasingcytoplasm volume or by treatment of embryos with anepigenetic modifier scriptaid.••Some developmentally important genes viz. Bcl-xl, Bax,Glut-1 and HSP 70.1 were found to be differentiallyexpressed in cloned and IVF-derived embryos.••Cloned buffalo embryos were produced by using somaticcells isolated from milk as donor cells.••Cardiomyocytes, which exhibited rhythmic beating, weregenerated from IVF goat ES cells.••An efficient protocol was established in goat for productionof embryonic stem cells from blastocysts produced byhandmade cloning.••Development of 3 buffalo embryonic stem cell linesachieved.••In vitro culture system capable of supporting long termself-renewal of buffalo embryonic stem cells was developed..Buffalo embryonic stem cells were characterized by abattery of markers such as SSEA-1, SSEA-3, SSEA-4,TRA-1-60, TRA-1-81, CD9, CD90, OCT4, SOX2,FOXD3, REX-1 and NUCLEOSTEMIN. Embryoidbodies were formed by spontaneous differentiation ofbuffalo embryonic stem cells, which contained cells from allthree germ layers i.e., ectoderm, mesoderm and endoderm,as confirmed by expression of markers specific to these celltypes.••Neurons and muscle cells from buffalo embryonic stemcells by directed differentiation were produced. Buffaloembryonic stem cells were produced using embryosproduced by Hand-guided cloning and parthenogenesis.Buffalo embryonic stem cells derived from blastocystsproduced by IVF, Handmade cloning and parthenogenesiswere shown to exhibit equivalency in terms of theirpluripotent marker expression. There exists a crosstalkbetween JAK-STAT and MAPK pathway and thatinhibition of LIF signaling caused inhibition ofpluripotency in buffalo embryonic stem cells. WNT3Aworks together with exogenous FGF-2 and LIF, resultingin proliferation of undifferentiated buffalo ES cells and thatWNT3A resulted in formation of scaffold like structureand inhibition of neural cell differentiation in differentiatedbuffalo ES cells. Buffalo (Bubalus bubalis) OCT4 orthologexpressed in buffalo embryonic stem cells and its promoterregion were cloned and characterized. Buffalo NANOGgene was cloned and characterized and alternativetranscription start cSites, splicing and polyadenylation wasexamined in buffalo embryonic stem cells.••During the course of in vitro culture, buffalo oocytesand embryos were found to suffer from DNA damage,which could be partly ameliorated by supplementationof culture media with cysteamine. Nitric oxide wasshown to be necessary for optimal embryo development inbuffalo. Lowering the O 2concentration during IVM, IVFand IVC from 20% to 5% or supplementation of IVM andIVC media with cysteamine, an antioxidant, was shown toimprove blastocyst yield, increase the expression of antiapoptoticgenes and decrease that of pro-apoptotic genes.••Single blastomere sexing of goat embryos was carried outby PCR amplification of the SRY gene.TheQuinquennium Golden2007-20127