Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
College of Natural and Mathematical Sciences, University of Maryland, Baltimore County, Baltimore, Maryland, USA.
Theriogenology. 2014 Jan 15;81(2):243-9. doi: 10.1016/j.theriogenology.2013.09.018. Epub 2013 Oct 21.
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.
犬脂肪间充质干细胞(ASCs)作为体细胞核移植(SCNT)的供体细胞具有很大的应用潜力。已有研究表明,不同的细胞培养条件可能会影响 SCNT 胚胎的早期发育。本研究旨在探讨两种不同的培养液(RCMEP、DMEM)是否会影响 ASCs 的基因表达、种间体细胞核移植(iSCNT)的后续发育以及克隆胚胎的基因表达。RCMEP 培养的细胞中 SOX2、NANOG、OCT4、DNMT1 和 MeCP2 的表达水平明显高于 DMEM 培养的细胞(P<0.05)。在 iSCNT 中,使用 DMEM 培养液培养细胞的胚胎发育至囊胚阶段的比例与 RCMEP 培养液培养的细胞相似(分别为 4.5%和 3.2%;P>0.05)。除了 DNMT1 以外,来源于 RCMEP 培养细胞的克隆囊胚和来源于 DMEM 培养细胞的克隆囊胚的所有转录本的表达水平相似。培养液诱导的 ASCs 基因表达的改变并未表现在由这些细胞产生的 iSCNT 胚胎中。虽然培养液可以诱导 ASCs 的基因表达发生改变,但这种供体细胞的改变不会影响 iSCNT 胚胎的发育能力或基因表达模式。