Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium
Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium
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DOI：
10.1073/pnas.0806136105
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发表时间：
2008-09-09
影响因子：
11.1
通讯作者：
Andrews, Peter W.
中科院分区：
文献类型：
--
作者：
Furue, Miho K.;Na, Jie;Andrews, Peter W.
开发人类胚胎干细胞（HESC）应用的一个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 最常维持在灭活的小鼠胚胎成纤维细胞饲养层中，培养基中补充有 FCS 或专有替代品（例如敲除血清替代品和 FGF-2）。这些未定义的培养条件阻碍了对控制 HESC 行为的机制的分析。我们现在开发了一种成分确定的无血清培养基 hESF9，用于在 I 型胶原基质上培养 HESC，无需饲养层。与其他报道的用于 HESC 培养的培养基相比，该培养基具有较低的渗透压（292 mosmol/升）、L-抗坏血酸-2-磷酸（0.1 μg/ml）和肝素。胰岛素、转铁蛋白、油酸结合白蛋白和 FGF-2 (10 ng/ml) 是唯一的蛋白质成分。此外，我们发现，如果同时存在肝素，HESC 在没有外源 FGF-2 的情况下也会增殖。然而，添加高达 10 ng/ml 的 FGF-2 可以增强它们的生长，尽管在肝素存在的情况下较高浓度是有害的。
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 mu g/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.