Co-culture of human embryonic stem cells with murine embryonic fibroblasts on microwell-patterned substrates
Co-culture of human embryonic stem cells with murine embryonic fibroblasts on microwell-patterned substrates
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DOI：
10.1016/j.biomaterials.2006.06.035
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发表时间：
2006-12-01
期刊：
影响因子：
14
通讯作者：
Langer, Robert
中科院分区：
文献类型：
--
作者：
Khademhosseini, Ali;Ferreira, Lino;Langer, Robert
人胚胎干（hES）细胞通常作为细胞簇在由有丝分裂失活的鼠胚胎成纤维细胞（MEF）形成的饲养层的顶部上培养以维持其未分化状态。这种通常用于扩增未分化hES细胞群体的共培养系统存在若干挑战，因为难以控制细胞簇大小。大的细胞团倾向于在边缘分化，不同大小的细胞团可能导致胚状体内的异质分化模式。在这项工作中，我们开发了一种新的方法来培养hES细胞与控制集群的大小和数量，通过合并微制造，和生物材料技术。制造聚合物微孔并用于控制与MEF共培养的hES细胞簇的大小和均匀性。结果表明，可以均匀培养hES细胞，同时保持其未分化状态，如干细胞标志物八聚体结合蛋白4（Oct-4）和碱性磷酸酶（ALP）的表达所证实的。此外，可以从微孔中回收这些簇，以产生用于分化实验的几乎均匀的细胞聚集体。(c)2006爱思唯尔有限公司保留所有权利。
Human embryonic stem (hES) cells are generally cultured as cell clusters on top of a feeder layer formed by mitotically inactivated murine embryonic fibroblasts (MEFs) to maintain their undifferentiated state. This co-culture system, which is typically used to expand the population of undifferentiated hES cells, presents several challenges since it is difficult to control cell cluster size. Large cell clusters tend to differentiate at the borders, and clusters with different sizes may lead to heterogeneous differentiation patterns within embryoid bodies. In this work, we develop a new approach to culture hES cells with controlled cluster size and number through merging microfabrication, and biomaterials technologies. Polymeric microwells were fabricated and used to control the size and uniformity of hES cell clusters in co-culture with MEFs. The results show that it is possible to culture hES cells homogeneously while keeping their undifferentiated state as confirmed by the expression of stem cell markers octamer binding protein 4 (Oct-4) and alkaline phosphatase (ALP). In addition, these clusters can be recovered from the microwells to generate nearly homogeneous cell aggregates for differentiation experiments. (c) 2006 Elsevier Ltd. All rights reserved.