Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V.
Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V.
复制标题
DOI：
10.1186/1471-2121-10-44
复制
发表时间：
2009-06-05
期刊：
影响因子：
--
通讯作者：
Ma W
中科院分区：
文献类型：
--
作者：
Tavakoli T;Xu X;Derby E;Serebryakova Y;Reid Y;Rao MS;Mattson MP;Ma W
胚胎干细胞的一个独特的和基本的属性是自我更新和分化成多个细胞谱系的能力。然而，独立衍生的人胚胎干细胞（hESC）之间的增殖和分化能力的可能差异并不为人所知，因为不充分的表征。为了解决这个问题，我们对以下两个方面进行了比较：1）在标准条件下维持未分化状态和自我更新的能力; 2）在分化胚状体时自发分化成三个初级胚生殖系的能力; 3）在NIH注册的三个hES细胞系I3（TE 03）、I6（TE 06）和BG 01 V之间进行定向神经分化反应。细胞系I3和I6具有正常的XX和正常的XY核型，而BG 01 V是衍生自核型正常的细胞系BG 01的具有异常核型的变异细胞系。使用免疫细胞化学、流式细胞术、qRT-PCR和MPSS，我们发现所有三种细胞系都活跃地增殖并表达相似的“干性”标志物，包括转录因子POU 5 F1/Oct 3/4和NANOG、糖脂SSEA 4和TRA-1-81以及碱性磷酸酶活性。所有细胞系在胚状体中分化为三种胚性生殖细胞系，在神经分化培养基中分化为神经细胞系。然而，在菌落形态，生长速度，BrdU掺入，和基因表达的相对丰度在未分化和分化状态的细胞系中观察到了深刻的变化。未分化的I3细胞生长明显较慢，但其分化潜力大于I6和BG 01 V。在相同的神经分化促进条件下，每个细胞系分化为神经祖细胞的能力不同。我们的比较分析提供了进一步的证据之间的相似性和差异性的自我更新，自发和定向分化的人胚胎干细胞系。这些差异可能与用于hESC系衍生的胚胎的性别、阶段、质量和遗传背景的遗传变异和/或在培养中传代期间获得的变化有关。
A unique and essential property of embryonic stem cells is the ability to self-renew and differentiate into multiple cell lineages. However, the possible differences in proliferation and differentiation capabilities among independently-derived human embryonic stem cells (hESCs) are not well known because of insufficient characterization. To address this question, a side-by-side comparison of 1) the ability to maintain an undifferentiated state and to self-renew under standard conditions; 2) the ability to spontaneously differentiate into three primary embryonic germ lineages in differentiating embryoid bodies; and 3) the responses to directed neural differentiation was made between three NIH registered hES cell lines I3 (TE03), I6 (TE06) and BG01V. Lines I3 and I6 possess normal XX and a normal XY karyotype while BG01V is a variant cell line with an abnormal karyotype derived from the karyotypically normal cell line BG01. Using immunocytochemistry, flow cytometry, qRT-PCR and MPSS, we found that all three cell lines actively proliferated and expressed similar "stemness" markers including transcription factors POU5F1/Oct3/4 and NANOG, glycolipids SSEA4 and TRA-1-81, and alkaline phosphatase activity. All cell lines differentiated into three embryonic germ lineages in embryoid bodies and into neural cell lineages when cultured in neural differentiation medium. However, a profound variation in colony morphology, growth rate, BrdU incorporation, and relative abundance of gene expression in undifferentiated and differentiated states of the cell lines was observed. Undifferentiated I3 cells grew significantly slower but their differentiation potential was greater than I6 and BG01V. Under the same neural differentiation-promoting conditions, the ability of each cell line to differentiate into neural progenitors varied. Our comparative analysis provides further evidence for similarities and differences between three hESC lines in self-renewal, and spontaneous and directed differentiation. These differences may be associated with inherited variation in the sex, stage, quality and genetic background of embryos used for hESC line derivation, and/or changes acquired during passaging in culture.
登录
查看更多内容
影响因子：
3.5
作者：
Enver, T;Soneji, S;Andrews, PW
通讯作者：
Andrews, PW
影响因子：
46.9
作者：
Reubinoff, BE;Itsykson, P;Ben-Hur, T
通讯作者：
Ben-Hur, T
影响因子：
5.2
作者：
Guhr, Anke;Kurtz, Andreas;Loeser, Peter
通讯作者：
Loeser, Peter
影响因子：
5.2
作者：
Ware, Carol B.;Nelson, Angelique M.;Blau, C. Anthony
通讯作者：
Blau, C. Anthony
影响因子：
5.2
作者：
Plaia, Todd W.;Josephson, Richard;Auerbach, Jonathan M.
通讯作者：
Auerbach, Jonathan M.