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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium
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Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium

机译：肝素在确定的无血清培养基中促进人类胚胎干细胞的生长

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摘要

A major limitation in developing applications for the use of human embryonic stem cells (HESCs) is our lack of knowledge of their responses to specific cues that control self-renewal, differentiation, and lineage selection. HESCs are most commonly maintained on inactivated mouse embryonic fibroblast feeders in medium supplemented with FCS, or proprietary replacements such as knockout serum-replacement together with FGF-2. These undefined culture conditions hamper analysis of the mechanisms that control HESC behavior. We have now developed a defined serum-free medium, hESF9, for the culture of HESCs on a type I-collagen substrate without feeders. In contrast to other reported media for the culture of HESCs, this medium has a lower osmolarity (292 mosmol/liter), L-ascorbic acid-2-phosphate (0.1 μg/ml), and heparin. Insulin, transferrin, albumin conjugated with oleic acid, and FGF-2 (10 ng/ml) were the only protein components. Further, we found that HESCs would proliferate in the absence of exogenous FGF-2 if heparin was also present. However, their growth was enhanced by the addition of FGF-2 up to 10 ng/ml although higher concentrations were deleterious in the presence of heparin.
机译：在开发使用人类胚胎干细胞（HESC）的应用程序中的主要限制是我们缺乏对它们对控制自我更新，分化和谱系选择的特定线索的反应的了解。 HESC最常在灭活的小鼠胚胎成纤维细胞饲养器上补充有FCS或专有替代品（例如与FGF-2一起进行基因敲除血清置换）维持。这些不确定的培养条件妨碍了对控制HESC行为的机制的分析。现在，我们已经开发出了定义的无血清培养基hESF9，用于在I型胶原底物上不加饲养器的情况下培养HESC。与其他报道的用于培养HESC的培养基相比，该培养基的渗透压较低（292 mosmol /升），L-抗坏血酸-2-磷酸（0.1μg/ ml）和肝素。胰岛素，转铁蛋白，与油酸结合的白蛋白和FGF-2（10 ng / ml）是唯一的蛋白质成分。此外，我们发现，如果还存在肝素，在没有外源性FGF-2的情况下HESCs会增殖。但是，通过添加高达10 ng / ml的FGF-2可以促进它们的生长，尽管在肝素存在下较高的浓度对人体有害。

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