Subpopulations of human embryonic stem cells with distinct tissue-specific fates can be selected from pluripotent cultures.
Subpopulations of human embryonic stem cells with distinct tissue-specific fates can be selected from pluripotent cultures.
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DOI：
10.1089/scd.2009.0012
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发表时间：
2009-12
影响因子：
4
通讯作者：
Bernstein HS
中科院分区：
文献类型：
--
作者：
King FW;Ritner C;Liszewski W;Kwan HC;Pedersen A;Leavitt AD;Bernstein HS
人胚胎干细胞（human embryonic stem cells，hESC）的定向分化在再生医学领域引起了广泛的关注。虽然已经基于特异性表面抗原的表达鉴定了多能培养物中的hESC亚群，但它们的意义和命运还没有很好地理解。为了确定这些亚群是否指示特定的组织命运或代表增殖培养物中的随机抗原分布，我们从组成型表达增强型绿色荧光蛋白（GFP）的增殖培养物中分离出CD 133+或CD 135 + hESC，并将其与GFP− hESC共培养。在培养物中传代后，对GFP+ hESC的CD 133或CD 135表达以及其他表面抗原（Tra-1-60、SSEA-4、FGFR-1）的持续性进行再分析，证明这两个亚群在连续传代中继续表达CD 133或CD 135，并且CD 133 + hESC也富集SSEA-4表达。在体外分化时，CD 133 +GFP+ hESC仅产生外胚层，如通过巢蛋白的表达所检测到的。在GFP+组织中未观察到代表内胚层（α-甲胎蛋白+）和中胚层（平滑肌肌动蛋白+）的组织。相反，针对CD 133的选择几乎只产生中胚层和内胚层。相比之下，CD 135 +GFP+ hESC产生的组织代表了所有三个胚胎胚层，并且几乎无法与CD 135 −衍生的组织区分开来。通过在畸胎瘤中的体内分化获得了类似的结果。这些数据证实存在其组织命运是预定的增殖hESC亚群，并挑战增殖hESC培养物内的所有细胞都是真正“多能性”的概念。这种共培养方法也将能够鉴定其他不同的hESC亚群，并且对这些亚群的选择应该证明在产生用于基于细胞的治疗的组织特异性试剂中是有价值的。
Directed differentiation of human embryonic stem cells (hESCs) has generated much interest in the field of regenerative medicine. While subpopulations of hESCs within pluripotent cultures have been identified based on expression of specific surface antigens, their significance and fates are not well understood. To determine whether such subpopulations indicate specific tissue fates or represent stochastic antigen distributions within proliferating cultures, we isolated CD133+ or CD135+ hESCs from proliferating cultures constitutively expressing enhanced green fluorescent protein (GFP), and co-cultured these with unselected GFP− hESCs. After passage in culture, GFP+ hESCs reanalyzed for the persistence of CD133 or CD135 expression, as well as other surface antigens (Tra-1-60, SSEA-4, FGFR-1), demonstrated that these two subpopulations continued to express CD133 or CD135 over serial passage, and that CD133+ hESCs were enriched for SSEA-4 expression as well. Upon differentiation in vitro, CD133+GFP+ hESCs gave rise solely to ectoderm, as detected by expression of nestin. Tissues representing endoderm (α-fetoprotein+) and mesoderm (smooth muscle actin+) were not seen among GFP+ tissues. In contrast, selection against CD133 gave rise almost exclusively to mesoderm and endoderm. In contrast, CD135+GFP+ hESCs gave rise to tissues representing all three embryonic germ layers, and were virtually indistinguishable from CD135−-derived tissues. Similar results were obtained by in vivo differentiation in teratomas. These data establish that subpopulations of proliferating hESCs whose tissue fate is predetermined exist, and challenge the notion that all cells within proliferating hESC cultures are truly “pluripotent.” This co-culture approach also will enable identification of other distinct hESC subpopulations, and selection for these should prove valuable in generating tissue-specific reagents for cell-based therapy.
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影响因子：
4
作者：
Epting, CL;López, JE;Bernstein, HS
通讯作者：
Bernstein, HS
影响因子：
11.2
作者：
Pfenninger, Cosima V.;Roschupkina, Teona;Nuber, Ulrike A.
通讯作者：
Nuber, Ulrike A.
影响因子：
4
作者：
Peh, Gary S. -L.;Lang, Richard J.;Hawes, Susan M.
通讯作者：
Hawes, Susan M.
影响因子：
5.3
作者：
Sawamoto, K;Nakao, N;Okano, H
通讯作者：
Okano, H
影响因子：
--
作者：
CUNHA, GR
通讯作者：
CUNHA, GR