Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions
Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions
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DOI：
10.1073/pnas.1114854108
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发表时间：
2011-11-15
影响因子：
11.1
通讯作者：
Jaenisch, Rudolf
中科院分区：
文献类型：
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作者：
Saha, Krishanu;Mei, Ying;Jaenisch, Rudolf
目前培养人多能干细胞的金标准需要使用细胞饲养层。在这里，我们开发了一个空间定义的文化系统的基础上，典型的细胞培养塑料的UV/臭氧辐射改性，以定义一个有利的表面环境，人类多能干细胞培养。表面的化学和几何优化使得能够控制来自完全解离的细胞的早期细胞聚集，如从细胞迁移的数值模型预测的，并且导致未分化细胞的细胞生长的显著增加。这些化学成分确定的无异种基质每表面积产生的细胞数量是含饲养层基质的三倍以上。此外，重编程和典型的基因靶向方案可以容易地在这些工程化表面上进行。这些基质为从患者组织样品中生产临床相关的无因子重编程细胞提供了一个有吸引力的细胞培养平台，并有助于定义用于疾病建模和细胞治疗应用的标准化放大友好方法。
The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.