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Effect of culture medium type on canine adipose-derived mesenchymal stem cells and developmental competence of interspecies cloned embryos.

机译:培养基类型对犬类脂肪间充质干细胞和种间克隆胚胎发育能力的影响。

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摘要

Canine adipose-derived mesenchymal stem cells (ASCs) are promising as donor cells for somatic cell nuclear transfer (SCNT). It has been suggested that different cell cultures possess different capacities to support pre-implantation development of SCNT embryos. The aim of this study is to investigate whether two culture medium (RCMEP, Dulbecco's modified Eagle's medium [DMEM]) affect gene expression of ASCs, subsequent development of interspecies SCNT (iSCNT) and gene expression of cloned embryos. The RCMEP-cultured cells contained significantly greater amounts of SOX2, NANOG, OCT4, DNMT1, and MeCP2 than DMEM-cultured cells (P<0.05). In iSCNT, the use of DMEM medium for culturing cells resulted in similar development to the blastocyst stage than those derived from RCMEP cultured cells (4.5% and 3.2%, respectively; P>0.05). The expression of all transcripts except for DNMT1 in cloned blastocysts from RCMEP cultured cells followed those of cloned blastocysts derived from DMEM cultured cells. The alteration of gene expression in ASCs by culture medium was not manifested in the iSCNT embryos derived from these cells. Although the culture medium can induce changes of gene expression by ASCs, such alterations in donor cells did not affect the developmental competence or gene expression patterns of iSCNT embryos.
机译:犬脂肪来源的间充质干细胞(ASCs)有望作为体细胞核移植(SCNT)的供体细胞。已经提出,不同的细胞培养物具有支持SCNT胚胎植入前发育的不同能力。这项研究的目的是调查两种培养基(RCMEP,Dulbecco改良的Eagle培养基[DMEM])是否会影响ASC的基因表达,种间SCNT(iSCNT)的后续发展以及克隆胚胎的基因表达。与DMEM培养的细胞相比,RCMEP培养的细胞含有大量的SOX2,NANOG,OCT4,DNMT1和MeCP2(P <0.05)。在iSCNT中,与从RCMEP培养的细胞衍生的细胞相比,使用DMEM培养基培养细胞的胚泡期发育相似(分别为4.5%和3.2%; P> 0.05)。在从RCMEP培养的细胞克隆的囊胚中,除DNMT1以外的所有转录本的表达都与从DMEM培养的细胞克隆的胚泡中的转录子相同。在这些细胞来源的iSCNT胚胎中没有发现培养基对ASCs基因表达的改变。尽管培养基可以通过ASC诱导基因表达的改变,但供体细胞中的这种改变并不影响iSCNT胚胎的发育能力或基因表达模式。

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