Department of Experimental Embryology, Institute of Genetics and Animal Biotechnology, Polish Academy of Sciences, Jastrzębiec,05-552 Magdalenka, Poland.
Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk & Medical University of Gdańsk, 80-307 Gdańsk, Poland.
Cells. 2021 Feb 25;10(3):490. doi: 10.3390/cells10030490.
The phenomenon of the reprogramming of terminally differentiated cells can be achieved by various means, like somatic cell nuclear transfer, cell fusion with a pluripotent cell, or the introduction of pluripotency genes. Here, we present the evidence that somatic cells can attain the expression of pluripotency markers after their introduction into early embryos. Mouse embryonic fibroblasts introduced between blastomeres of cleaving embryos, within two days of in vitro culture, express transcription factors specific to blastocyst lineages, including pluripotency factors. Analysis of donor tissue marker DNA has revealed that the progeny of introduced cells are found in somatic tissues of foetuses and adult chimaeras, providing evidence for cell reprogramming. Analysis of ploidy has shown that in the chimaeras, the progeny of introduced cells are either diploid or tetraploid, the latter indicating cell fusion. The presence of donor DNA in diploid cells from chimaeric embryos proved that the non-fused progeny of introduced fibroblasts persisted in chimaeras, which is evidence of reprogramming by embryonic niche. When adult somatic (cumulus) cells were introduced into early cleavage embryos, the extent of integration was limited and only cell fusion-mediated reprogramming was observed. These results show that both cell fusion and cell interactions with the embryonic niche reprogrammed somatic cells towards pluripotency.
通过各种方法可以实现终末分化细胞的重编程,如体细胞核移植、与多能细胞融合,或导入多能性基因。在这里,我们提供了证据表明,体细胞在被导入早期胚胎后可以获得多能性标记物的表达。将体外培养两天内的分裂胚胎的卵裂球之间的小鼠胚胎成纤维细胞导入其中,可表达囊胚谱系特有的转录因子,包括多能性因子。对供体细胞标记物 DNA 的分析表明,导入细胞的后代存在于胎儿和成体嵌合体的体细胞组织中,这为细胞重编程提供了证据。对倍性的分析表明,在嵌合体中,导入细胞的后代要么是二倍体,要么是四倍体,后者表明细胞融合。来自嵌合体胚胎的二倍体细胞中存在供体 DNA,证明了在嵌合体中,导入的成纤维细胞的未融合后代持续存在,这是胚胎生态位重编程的证据。当将成年体(卵丘)细胞导入早期分裂胚胎时,整合的程度有限,仅观察到细胞融合介导的重编程。这些结果表明,细胞融合和与胚胎生态位的细胞相互作用都可以将体细胞重编程为多能性。