An intracytoplasmic injection of deionized bovine serum albumin immediately after somatic cell
nuclear transfer enhances full-term development of cloned mouse embryos |
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Authors: | Yuuki ISAJI Koki YOSHIDA Hiroshi IMAI Masayasu YAMADA |
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Institution: | 1)Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan |
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Abstract: | In mouse somatic cell nuclear transfer (SCNT), polyvinylpyrrolidone (PVP) is typically included in the
nuclear donor injection medium. However, the cytotoxicity of PVP, which is injected into the cytoplasm of
oocytes, has recently become a cause of concern. In the present study, we determined whether bovine serum
albumin deionized with an ion-exchange resin treatment (d-BSA) was applicable to the nuclear donor injection
medium in SCNT as an alternative to PVP. The results obtained showed that d-BSA introduced into the cytoplasm
of an enucleated oocyte together with a donor nucleus significantly enhanced the rate of in
vitro development of cloned embryos to the blastocyst stage compared with that of a conventional
nuclear injection with PVP in SCNT. We also defined the enhancing effects of d-BSA on the blastocyst formation
rate when d-BSA was injected into the cytoplasm of oocytes reconstructed using the fusion method with a
hemagglutinating virus of Japan envelope before oocyte activation. Furthermore, immunofluorescence experiments
revealed that the injected d-BSA increased the acetylation levels of histone H3 lysine 9 and histone H4 lysine
12 in cloned pronuclear (PN) and 2-cell embryos. The injection of d-BSA before oocyte activation also
increased the production of cloned mouse offspring. These results suggested that intracytoplasmic injection of
d-BSA into SCNT oocytes before oocyte activation was beneficial for enhancing the in vitro
and in vivo development of mouse cloned embryos through epigenetic modifications to nuclear
reprogramming. |
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Keywords: | Deionized bovine serum albumin (d-BSA) Full-term development Histone acetylation Mouse Somatic cell nuclear transfer (SCNT) |
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