Publication: Somatic cell cloning in Buffalo (Bubalus bubalis): Effects of interspecies cytoplasmic recipients and activation procedures
Issued Date
2001-01-01
Resource Type
ISSN
15362302
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2-s2.0-0035704630
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Mahidol University
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SCOPUS
Bibliographic Citation
Cloning and Stem Cells. Vol.3, No.3 (2001), 97-104
Suggested Citation
Yindee Kitiyanant, Jumnian Saikhun, Busabun Chaisalee, Kenneth L. White, Kanok Pavasuthipaisit Somatic cell cloning in Buffalo (Bubalus bubalis): Effects of interspecies cytoplasmic recipients and activation procedures. Cloning and Stem Cells. Vol.3, No.3 (2001), 97-104. doi:10.1089/153623001753205052 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/26486
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Title
Somatic cell cloning in Buffalo (Bubalus bubalis): Effects of interspecies cytoplasmic recipients and activation procedures
Abstract
Successful nuclear transfer (NT) of somatic cell nuclei from various mammalian species to enucleated bovine oocytes provides a universal cytoplast for NT in endangered or extinct species. Buffalo fetal fibroblasts were isolated from a day 40 fetus and were synchronized in presumptive G0by serum deprivation. Buffalo and bovine oocytes from abattoir ovaries were matured in vitro and enucleated at 22 h. In the first experiment, we compared the ability of buffalo and bovine oocyte cytoplasm to support in vitro development of NT embryos produced by buffalo fetal fibroblasts as donor nuclei. There were no significant differences (p > 0.05) between the NT embryos derived from buffalo and bovine oocytes, in fusion (74% versus 71%) and cleavage (77% versus 75%) rates, respectively. No significant differences were also observed in blastocyst development (39% versus 33%) and the mean cell numbers of day 7 cloned blastocysts (88.5 ± 25.7 versus 51.7 ± 5.4). In the second experiment, we evaluated the effects of activation with calcium ionophore A23187 on development of NT embryos after electrical fusion. A significantly higher (p < 0.05) percentage of blastocyst development was observed in the NT embryos activated by calcium ionophore and 6-DMAP when compared with 6-DMAP alone (33% versus 17%). The results indicate that the somatic nuclei from buffalo can be reprogrammed after transfer to enucleated bovine oocytes, resulting in the production of cloned buffalo blastocysts similar to those transferred into buffalo oocytes. Calcium ionophore used in conjunction with 6-DMAP effectively induces NT embryo development.