Expansion of pluripotent human embryonic stem cells on human feeders
Expansion of pluripotent human embryonic stem cells on human feeders
复制标题
DOI：
10.1002/bit.20247
复制
发表时间：
2004-11-05
影响因子：
3.8
通讯作者：
Oh, SKW
中科院分区：
文献类型：
--
作者：
Choo, ABH;Padmanabhan, J;Oh, SKW
人类胚胎干细胞（HES）由于能够分化为任何细胞类型，因此在再生医学方面具有巨大的潜力。然而，一个限制是 HES 细胞需要饲养层才能保持未分化。通常使用小鼠胚胎成纤维细胞。然而，对于治疗应用，小鼠细胞的污染可能被认为是不可接受的。在这项研究中，我们评估了三种市售的人类包皮饲养 (HF) 系，以评估它们在补充有血清或血清替代品的培养基中支持 HES 细胞生长的能力。 HES 细胞在添加血清替代物的培养基中进行 HF 培养 > 30 代。它们保持未分化，保持正常核型，并且多能标记 Oct-4、SOX-2、SSEA-4、GCTM-2、Tra-1-60、Tra-1-81 和碱性磷酸酶继续呈阳性。在体内，HES 细胞在 SCID 小鼠模型中形成代表三个胚胎胚层的畸胎瘤。相比之下，在补充血清的培养基中用 HF 培养的 HES 细胞在传代 3 代后分化。从形态上看，细胞变成囊性，胞内 Oct-4 缺失。我们已经成功地在三个人类饲养系上适应和培养未分化的 HES 细胞超过 30 代。除了培养基中的血清外，没有观察到任何困难。这项研究揭示了用于 HES 细胞研究和潜在治疗应用的安全且易于获取的饲养源。 (C) 2004 年 Wiley 期刊公司。
Human embryonic stem cells (HES) hold great potential for regenerative medicine because of their ability to differentiate to any cell type. However, a limitation is that HES cells require a feeder layer to stay undifferentiated. Routinely, mouse embryonic fibroblast is used. However, for therapeutic applications, contamination with mouse cells may be considered unacceptable. In this study, we evaluated three commercially available human foreskin feeder (HF) lines for their ability to support HES cell growth in media supplemented with serum or serum replacer. HES cells on HF in serum replacer-supplemented media were cultured for >30 passages. They remained undifferentiated, maintained a normal karyotype, and continued to be positive for the pluripotent markers Oct-4, SOX-2, SSEA-4, GCTM-2, Tra-1-60, Tra-1-81, and alkaline phosphatase. In vivo, HES cells formed teratomas in SCID mouse models that represent the three embryonic germ layers. In contrast, HES cells cultured on HF in serum-supplemented media differentiated after three passages. Morphologically, the cells became cystic with a loss of intracellular Oct-4. We have successfully adapted and cultured undifferentiated HES cells on three human feeder lines for >30 passages. No difficulties were observed with the exception of serum in the media. This study reveals a safe and accessible source for feeders for HES cell research and potential therapeutic applications. (C) 2004 Wiley Periodicals, Inc.