Bcl-xL enhances single-cell survival and expansion of human embryonic stem cells without affecting self-renewal.
Bcl-xL enhances single-cell survival and expansion of human embryonic stem cells without affecting self-renewal.
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BCl-XL增强了人类干细胞的单细胞存活和扩展，而不会影响自我更新。
DOI：
10.1016/j.scr.2011.08.002
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发表时间：
2012-01
影响因子：
1.2
通讯作者：
Wang, Zack Z.
中科院分区：
文献类型：
--
作者：
Bai, Hao;Chen, Kang;Gao, Yong-Xing;Arzigian, Melanie;Xie, Yin-Liang;Malcosky, Christopher;Yang, Yong-Guang;Wu, Wen-Shu;Wang, Zack Z.
人类胚胎干细胞(HESCs)和诱导多能干细胞(IPS)的强劲扩增和遗传操作受到酶解离成单细胞后细胞存活率较低的限制。虽然抑制凋亡与hESCs的单细胞存活有关，但抑制凋亡在hESC存活中的保护作用尚未阐明。BCL-XL是几种抗细胞凋亡蛋白之一，属于Bcl2蛋白家族的成员。利用诱导系统，我们在hESCs中异位表达了bclxl基因，并在单细胞悬浮培养中发现hESC集落显著增加。在hESCs中过表达bclxl减少了caspase-3+细胞的凋亡，提示hESCs的凋亡减弱。在不改变多能基因表达动力学的情况下，Bclxl高表达的hESCs经单细胞分离后，在体外生成类胚体(EBS)的效率和体内畸胎瘤的形成显著增加。有趣的是，来自集群培养的hESC克隆的数量和大小不受Bclxl过表达的影响。在hESCs中，细胞外基质和黏附分子的几个基因在没有单细胞分离的情况下被Bclxl上调，提示Bclxl调节黏附分子的表达不依赖细胞解离。此外，Bclxl还下调了Fas和几种肿瘤坏死因子信号介质的基因表达。这些数据支持这样一种模型，即在不改变hESC自我更新的情况下，Bcl-xL促进细胞存活并提高hESCs的克隆效率，其机制如下：1)抑制细胞凋亡；2)上调黏附分子以促进细胞与细胞或细胞与基质的相互作用。
Robust expansion and genetic manipulation of human embryonic stem cells (hESCs) and induced-pluripotent stem (iPS) cells are limited by poor cell survival after enzymatic dissociation into single cells. Although inhibition of apoptosis is implicated for the single-cell survival of hESCs, the protective role of attenuation of apoptosis in hESC survival has not been elucidated. Bcl-xL is one of several anti-apoptotic proteins, which are members of the Bcl-2 family of proteins. Using an inducible system, we ectopically expressed Bcl-xL gene in hESCs, and found a significant increase of hESC colonies in the single-cell suspension cultures. Overexpression of Bcl-xL in hESCs decreased apoptotic caspase-3+ cells, suggesting attenuation of apoptosis in hESCs. Without altering the kinetics of pluripotent gene expression, the efficiency to generate embryoid bodies (EBs) in vitro and the formation of teratoma in vivo were significantly increased in Bcl-xL-overexpressing hESCs after single-cell dissociation. Interestingly, the number and size of hESC colonies from cluster cultures was not affected by Bcl-xL overexpression. Several genes of extracellular matrix and adhesion molecules were upregulated by Bcl-xL in hESCs without single-cell dissociation, suggesting that Bcl-xL regulates adhesion molecular expression independent of cell dissociation. In addition, the gene expression of FAS and several TNF signaling mediators were downregulated by Bcl-xL. These data support a model in which Bcl-xL promotes cell survival and increases cloning efficiency of dissociated hESCs without altering hESC self-renewal by i) attenuation of apoptosis, and ii) upregulation of adhesion molecules to facilitate cell-cell or cell-matrix interactions.
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影响因子：
4
作者：
Bai, Hao;Gao, Yongxing;Arzigian, Melanie;Wojchowski, Don M.;Wu, Wen-shu;Wang, Zack Z.
通讯作者：
Wang, Zack Z.
克隆衍生的人类胚胎干细胞系在长期培养过程中保持多能性和增殖潜力
影响因子：
2.7
作者：
Amit, M;Carpenter, MK;Thomson, JA
通讯作者：
Thomson, JA
DOI：
10.1073/pnas.1019047108
发表时间：
2011-02-22
影响因子：
11.1
作者：
Ardehali, Reza;Inlay, Matthew A.;Weissman, Irving L.
通讯作者：
Weissman, Irving L.
影响因子：
21.3
作者：
Feng, B;Yao, PM;Tabas, I
通讯作者：
Tabas, I
基质细胞衍生的 Factor-1/CXCR4 信号传导在体外改变人胚胎干细胞衍生的内皮细胞的毛细血管样组织
影响因子：
5.2
作者：
Chen, Tong;Bai, Hao;Wang, Zack Z.
通讯作者：
Wang, Zack Z.