Time-lapse analysis of human embryonic stem cells reveals multiple bottlenecks restricting colony formation and their relief upon culture adaptation.
Time-lapse analysis of human embryonic stem cells reveals multiple bottlenecks restricting colony formation and their relief upon culture adaptation.
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DOI：
10.1016/j.stemcr.2014.05.006
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发表时间：
2014-07-08
影响因子：
5.9
通讯作者：
Andrews, Peter W.
中科院分区：
文献类型：
--
作者：
Barbaric, Ivana;Biga, Veronica;Gokhale, Paul J.;Jones, Mark;Stavish, Dylan;Glen, Adam;Coca, Daniel;Andrews, Peter W.
使用延时成像，我们已经确定了一系列限制早期传代人胚胎干细胞（hESC）生长的瓶颈，并通过长时间培养选择的核型异常变体来缓解。只有少数核型正常的细胞在接种后分裂，这些细胞主要是接种时处于细胞周期后期的细胞。此外，子细胞在分裂后表现出持续的细胞死亡模式，因此很少形成长期增殖的集落。这些集落形成细胞显示出不同的细胞运动模式。增加细胞密度增强细胞运动，促进细胞：细胞接触，这导致分裂细胞的比例增加，并提高正常人胚胎干细胞接种后的存活率。相比之下，大多数核型异常的细胞在平板上重新进入细胞周期，并产生健康的后代，而不需要细胞：细胞接触和独立的运动模式。核型正常的hESC的生长受到多个瓶颈的限制。瓶颈包括初始存活、进入有丝分裂和有丝分裂后的死亡。正常hESC中的特权克隆形成状态的特征在于不同的运动性。变体适应细胞和ROCK抑制缓解了所鉴定的瓶颈Andrews，Coca，和他的同事们使用单人类胚胎干细胞（hESC）的延时成像，以确定它们的行为倾向于限制生长的方面，从而成为提供选择优势的突变的目标。他们确定了一系列限制早期传代hESC生长的瓶颈，并且通过长期培养选择的核型异常变体来缓解。
Using time-lapse imaging, we have identified a series of bottlenecks that restrict growth of early-passage human embryonic stem cells (hESCs) and that are relieved by karyotypically abnormal variants that are selected by prolonged culture. Only a minority of karyotypically normal cells divided after plating, and these were mainly cells in the later stages of cell cycle at the time of plating. Furthermore, the daughter cells showed a continued pattern of cell death after division, so that few formed long-term proliferating colonies. These colony-forming cells showed distinct patterns of cell movement. Increasing cell density enhanced cell movement facilitating cell:cell contact, which resulted in increased proportion of dividing cells and improved survival postplating of normal hESCs. In contrast, most of the karyotypically abnormal cells reentered the cell cycle on plating and gave rise to healthy progeny, without the need for cell:cell contacts and independent of their motility patterns. The growth of karyotypically normal hESCs is limited by multiple bottlenecks Bottlenecks include initial survival, entry into mitosis, and death following mitosis A privileged clonogenic state in normal hESCs is characterized by distinct motility Variant adapted cells and ROCK inhibition alleviate identified bottlenecks Andrews, Coca, and colleagues use time-lapse imaging of single human embryonic stem cells (hESCs) to identify aspects of their behavior that tend to restrict growth and so would be targets for mutations that provide selective advantage. They identify a series of bottlenecks that restricts growth of early-passage hESCs and that is relieved by karyotypically abnormal variants selected by prolonged culture.
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影响因子：
5.6
作者：
Becker, Klaus A.;Ghule, Prachi N.;Stein, Gary S.
通讯作者：
Stein, Gary S.
影响因子：
20.3
作者：
Joshi, Ila;Minter, Lisa M.;Osborne, Barbara A.
通讯作者：
Osborne, Barbara A.
影响因子：
3.5
作者：
Enver, T;Soneji, S;Andrews, PW
通讯作者：
Andrews, PW
影响因子：
5.3
作者：
Rand, MD;Grimm, LM;Aster, JC
通讯作者：
Aster, JC
Notch 激活静止心肌细胞的细胞周期重入和进展。
DOI：
10.1083/jcb.200806104
发表时间：
2008-10-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Campa VM;Gutiérrez-Lanza R;Cerignoli F;Díaz-Trelles R;Nelson B;Tsuji T;Barcova M;Jiang W;Mercola M
通讯作者：
Mercola M