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首页> 外文期刊>Journal of Biotechnology >Adaptation of human embryonic stem cells to feeder-free and matrix-free culture conditions directly on plastic surfaces
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Adaptation of human embryonic stem cells to feeder-free and matrix-free culture conditions directly on plastic surfaces

机译：直接在塑料表面上使人胚胎干细胞适应无饲养者和无基质的培养条件

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Previous studies have shown that cultivation of undifferentiated human embryonic stem (hES) cells requires human fibroblasts (hF) or mouse embryonic fibroblast (mEF) feeders or a coating matrix such as laminin, fibronectin or Matrigeltrade mark in combination with mEF or hF conditioned medium. We here demonstrate a successful feeder-free and matrix-free culture system in which undifferentiated hES cells can be cultured directly on plastic surfaces without any supportive coating, in a hF conditioned medium. The hES cells cultured directly on plastic surfaces grow as colonies with morphology very similar to cells cultured on Matrigel(TM). Two hES cell lines SA167 and AS034.1 were adapted to matrix-free growth (MFG) and have so far been cultured up to 43 passages and cryopreserved successfully. The lines maintained a normal karyotype and expressed the expected marker profile of undifferentiated hES cells for Oct-4, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and SSEA-1. The hES cells formed teratomas in SCID mice and differentiated in vitro into derivates of all three germ layers. Thus, the MFG-adapted hES cells appear to retain pluripotency and to remain undifferentiated. The present culture system has a clear potential to be scaleable up to a manufacturing level and become the preferred culture system for various applications such as cell therapy and toxicity testing.
机译：以前的研究表明，未分化的人类胚胎干细胞（hES）的培养需要人类成纤维细胞（hF）或小鼠胚胎成纤维细胞（mEF）饲养者，或者将层粘连蛋白，纤连蛋白或Matrigeltrade标记等涂层基质与mEF或hF条件培养基结合使用。我们在这里展示了一个成功的无饲养者和无基质的培养系统，其中未分化的hES细胞可以在hF条件培养基中直接培养在塑料表面上，而无需任何支持性涂层。直接在塑料表面上培养的hES细胞以具有类似于在Matrigel™上培养的细胞的形态的菌落生长。两种hES细胞系SA167和AS034.1适用于无基质生长（MFG），到目前为止已培养了多达43代，并成功冷冻保存。该品系保持正常的核型，并表达了Oct-4，SSEA-3，SSEA-4，TRA-1-60，TRA-1-81和SSEA-1的未分化hES细胞的预期标记谱。 hES细胞在SCID小鼠中形成畸胎瘤，并在体外分化为所有三个胚层的衍生物。因此，适应MFG的hES细胞似乎保留了多能性并且保持未分化。本培养系统具有明显的潜力，可以扩大到制造水平，并成为各种应用如细胞疗法和毒性测试的优选培养系统。

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