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Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar

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20141
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Subject: Plant Physiology ×
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    Studies On donor cells for production of cloned embryos and their characterization In buffalo (bubalus bubalis)
    (LUVAS, 2014) Shah, Fozia; Gupta, Meenakshi
    Studies were carried out to establish bubaline fibroblast cell cultures and use them as nuclear donor for production of handmade cloned embryos, to compare the fresh and frozen donor cells for cloned embryo production, and analyze the expression of key cloning associated genes (GNAI2 /DNMT1 and BAX) in cloned couplets. Primary fibroblast cell cultures from newborn buffalo calf ear pinna were taken as a source of donor karyoplasts and after trypsinization were subjected to cryopreservation (-196⁰C). These were used as frozen cell donors for further study in the Handmade cloned embryos. mRNA was extracted from the fresh and frozen derived embryos. The results revealed that there were non significant differences in the cleavage rates of the cloned embryos derived from three different cell culture confluences (60-70%, 70-80% and 80–90%) but there were significant differences in the blastocyst formation rate. The highest blastocyst formation rate was observed at 80- 90% cell culture confluences among the three confluences. The in vitro developmental potential of cloned embryos derived from fresh fibroblast cells was higher than that of frozen fibroblast derived embryos in terms of blastocyst rate. Non significant differences were observed in the mRNA transcripts of the genes of the embryos derived from fresh and frozen fibroblast cells. Handmade cloned buffalo embryos derived from frozen fibroblast reveals expression of developmentally important genes GNAI2, DNMT1 and BAX at levels similar to fresh fibroblast derived embryos.