Comparative study of mouse and human feeder cells for human embryonic stem cells
Comparative study of mouse and human feeder cells for human embryonic stem cells
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DOI：
10.1387/ijdb.082590le
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发表时间：
2008-01-01
影响因子：
0.7
通讯作者：
Dvorak, Petr
中科院分区：
文献类型：
--
作者：
Eiselleova, Livia;Peterkova, Iveta;Dvorak, Petr
为了提高人胚胎干细胞（hESCs）的附着性并为其提供干细胞支持因子，采用了多种类型的饲养细胞进行培养。然而，饲养细胞支持未分化hESCs生长的能力不同。在这里，我们比较了在标准hESC培养过程中，五种人类包皮成纤维细胞和小鼠原代胚胎成纤维细胞中四种成熟的hESC多能性和/或分化调节因子的表达和分泌。我们发现人和小鼠饲养细胞分泌的TGF β 1水平相当。然而，小鼠饲养细胞比人类饲养细胞分泌更多的激活素A。相反，由人饲养细胞产生的FGF-2在小鼠饲养细胞的培养基中无法检测到。虽然BMP-4二聚体存在于所有饲养细胞中，但BMP-4的含量在所有饲养细胞类型的培养基中都处于可检测水平。TGF β 1、激活素A和FGF-2的产生在人源性饲养细胞系中有很大差异。我们评估了低产量和高产产量的人饲养细胞以及小鼠饲养细胞支持hESCs未分化生长的能力。我们发现，在表达SSEA3（一种未分化细胞标记物）的人饲养细胞类型上维持的hESCs比例明显较低。此外，激活素a的加入可以部分挽救SSEA3的表达，从而挽救多能hESC细胞室。综上所述，这些结果表明，饲喂层促进hESCs未分化生长的能力可归因于其特征性生长因子的产生。
Various types of feeder cells have been adopted for the culture of human embryonic stem cells (hESCs) to improve their attachment and provide them with stemness-supporting factors. However, feeder cells differ in their capacity to support the growth of undifferentiated hESCs. Here, we compared the expression and secretion of four well-established regulators of hESC pluripotency and/or differentiation among five lines of human foreskin fibroblasts and primary mouse embryonic fibroblasts throughout a standard hESC culture procedure. We found that human and mouse feeder cells secreted comparable levels of TGF beta 1. However, mouse feeder cells secreted larger quantities of activin A than human feeder cells. Conversely, FGF-2, which was produced by human feeder cells, could not be detected in culture media from mouse feeder cells. The quantity of BMP-4 was at about the level of detectability in media from all feeder cell types, although BMP-4 dimers were present in all feeder cells. Production of TGF beta 1, activin A, and FGF-2 varied considerably among the human-derived feeder cell lines. Low- and high-producing human feeder cells as well as mouse feeder cells were evaluated for their ability to support the undifferentiated growth of hESCs. We found that a significantly lower proportion of hESCs maintained on human feeder cell types expressed SSEA3, an undifferentiated cell marker. Moreover, SSEA3 expression and thus the pluripotent hESC compartment could be partially rescued by addition of activin A. Cumulatively, these results suggest that the ability of a feeder layer to promote the undifferentiated growth of hESCs is attributable to its characteristic growth factor production.