Khosla S, Dean W, Brown D, Reik W, Feil R
Laboratory of Developmental Genetics and Imprinting and Laboratory of Computational Neuroscience, The Babraham Institute, Babraham, Cambridge CB2 4AT, United Kingdom.
Biol Reprod. 2001 Mar;64(3):918-26. doi: 10.1095/biolreprod64.3.918.
Culture of preimplantation mammalian embryos and cells can influence their subsequent growth and differentiation. Previously, we reported that culture of mouse embryonic stem cells is associated with deregulation of genomic imprinting and affects the potential for these cells to develop into normal fetuses. The purpose of our current study was to determine whether culture of preimplantation mouse embryos in a chemically defined medium (M16) with or without fetal calf serum (FCS) can affect their subsequent development and imprinted gene expression. Only one third of the blastocysts that had been cultured from two-cell embryos in M16 medium complemented with FCS developed into viable Day 14 fetuses after transfer into recipients. These M16 + FCS fetuses were reduced in weight as compared with controls and M16 fetuses and had decreased expression of the imprinted H19 and insulin-like growth factor 2 genes associated with a gain of DNA methylation at an imprinting control region upstream of H19. They also displayed increased expression of the imprinted gene Grb10. The growth factor receptor binding gene Grb7, in contrast, was strongly reduced in its expression in most of the M16 + FCS fetuses. No alterations were detected for the imprinted gene MEST: Preimplantation culture in the presence of serum can influence the regulation of multiple growth-related imprinted genes, thus leading to aberrant fetal growth and development.
植入前哺乳动物胚胎和细胞的培养会影响其随后的生长和分化。此前,我们报道过小鼠胚胎干细胞的培养与基因组印记调控异常有关,并影响这些细胞发育成正常胎儿的潜力。我们当前研究的目的是确定在含有或不含胎牛血清(FCS)的化学限定培养基(M16)中培养植入前小鼠胚胎是否会影响其随后的发育和印记基因表达。从二细胞胚胎在添加了FCS的M16培养基中培养得到的囊胚,只有三分之一在移植到受体后发育成了存活的第14天胎儿。与对照组和M16培养基培养的胎儿相比,这些M16 + FCS培养基培养的胎儿体重减轻,印记基因H19和胰岛素样生长因子2的表达降低,这与H19上游印记控制区域的DNA甲基化增加有关。它们还表现出印记基因Grb10的表达增加。相比之下,生长因子受体结合基因Grb7在大多数M16 + FCS培养基培养的胎儿中的表达大幅降低。未检测到印记基因MEST有改变:在有血清存在的情况下进行植入前培养会影响多个与生长相关的印记基因的调控,从而导致胎儿生长发育异常。