非休眠期成年兔骨髓基质干细胞和脂肪基质干细胞用于核移植构建重组胚

目的研究非休眠期成年兔骨髓基质干细胞和脂肪基质干细胞作为供体,以体外成熟去核兔卵母细胞作为受体进行核移植构建重组胚胎的可行性及囊胚形成率. 方法首先利用新西兰兔骨髓和脂肪为来源分别培养骨髓基质干细胞和脂肪基质干细胞,然后用他们作为核供体,采用卵细胞胞质内直接注射法构建重组胚胎,7 d后比较囊胚形成率. 结果核移植操作后卵细胞存活率达80.0%,骨髓基质干细胞和脂肪基质干细胞作为供体重组胚胎囊胚形成率分别为88/522 (16.9%)和73/464 (15.7%), (P>0.05, Yates corrected). 结论处于非休眠期的成年兔骨髓基质干细胞和脂肪基质干细胞移植到去核卵母细胞后能够进行早期胚胎发育,两者囊胚形成率无明显差别.

Production of blastocysts by transfer of nuclei from nonquiescent adult rabbits bone mesenchymal stem cells and adipose stromal stem cells

AIM To study the isolation and in vitro culture of rabbit's bone mesenchymal stem cells (BMSCs) and adipose stromal stem cells (ASSCs) and the use of both BMSCs and ASSCs cells in nuclear transfer (NT) to produce totipotent blastocysts. METHODS Both cell lines were derived from New Zealand rabbits and used as donor cells. The cell lines were cultured as a low cell density suspension in the Dulbecco's modified Eagle's medium (DMEM) and 150 Ml*L-1 fetal calf serum (FCS) for 7 d before use in nuclear transfer. Direct intracystplasm microinjection techniques were used to reconstruct one-cell embryos. Sire differences were noted in the ability of both BMSCs and ASSCs to form blastocysts. RESULTS A total of 986 nuclear transfer clones were made by fusing both BMSCs and ASSCs cells nuclei into enucleated oocytes. After culture of the clones for 7 d in vitro, 88 (16.9%) and 73 (15.7%) of those fused with BMSCs and ASSCs cells nuclei became blastocysts (P>0.05, Yates corrected). CONCLUSION BMSCs and ASSCs can be used as the NT donor cells to form blastocysts. There is no significant difference in blastocyst development rate (BDR) between them.