Guo Xin, Qi Zheng-Yu, Qin Jie, Cui Guang-Hui, Gui Yao-Ting, Cai Zhi-Ming
Key Laboratory of Male Reproduction and Genetics of Guangdong Province, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, China.
Zhonghua Nan Ke Xue. 2009 Nov;15(11):967-73.
Interactions of cells with the extracellular matrix (ECM) are essential for cell differentiation. The authors sought to determine the roles of different ECMs in the expressions of germ cell differentiation associated genes after mouse embryonic stem cells (mESCs) differentiated into embryoid bodies (EBs).
EBs derived from mESCs were maintained in suspension for 3 days and then cultured on the plates coated with various ECMs, including fibronectin (F), laminin (L), matrigel (M), collagen (C) and nonadhensive agarose (A), respectively, for 1, 2, 3 or 4 days, followed by evaluation of the expressions of the genes associated with germ cell differentiation by RT-PCR.
The EBs of the F and L groups exhibited facilitated adherent differentiation. The expressions of the Blimp-1, Stella, Mvh and Stra8 genes were increased gradually in the F and L but not obviously in the M and C groups. The overall gene expressions were low in the A group, but high and then gradually decreased in the blank control group. Endogenous fibronectin, laminin and integrin beta1 were obviously expressed in the L and control groups.
Laminin /integrin beta1 signaling may play a role in regulating the differentiation of mESCs into primordial germ cells (PGCs). Exogenous laminin can facilitate the differentiation of mESC-derived EBs into PGCs by acting on the integrin beta1 subunit, while exogenous fibronectin may be involved in the regulation of the differentiation through other integrin subunit. Endogenous laminin and fibronectin secreted by EBs may also facilitate cell differentiation in the absence of exogenous ECMs.
细胞与细胞外基质(ECM)的相互作用对细胞分化至关重要。作者试图确定在小鼠胚胎干细胞(mESCs)分化为胚状体(EBs)后,不同的细胞外基质在生殖细胞分化相关基因表达中的作用。
将源自mESCs的EBs悬浮培养3天,然后分别接种于包被有不同细胞外基质的培养板上,这些细胞外基质包括纤连蛋白(F)、层粘连蛋白(L)、基质胶(M)、胶原蛋白(C)和非黏附性琼脂糖(A),培养1、2、3或4天,随后通过逆转录聚合酶链反应(RT-PCR)评估生殖细胞分化相关基因的表达。
F组和L组的EBs表现出促进的贴壁分化。Blimp-1、Stella、Mvh和Stra8基因的表达在F组和L组中逐渐增加,而在M组和C组中不明显。A组的整体基因表达较低,而空白对照组的基因表达较高,然后逐渐下降。L组和对照组中内源性纤连蛋白、层粘连蛋白和整合素β1明显表达。
层粘连蛋白/整合素β1信号通路可能在调节mESCs向原始生殖细胞(PGCs)分化中发挥作用。外源性层粘连蛋白可通过作用于整合素β1亚基促进mESC来源的EBs分化为PGCs,而外源性纤连蛋白可能通过其他整合素亚基参与分化的调节。EBs分泌的内源性层粘连蛋白和纤连蛋白在没有外源性细胞外基质的情况下也可能促进细胞分化。