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A Novel Feeder-Free Culture System for Human Pluripotent Stem Cell Culture and Induced Pluripotent Stem Cell Derivation

机译:人多能干细胞培养和诱导多能干细胞衍生的新型无饲养员培养系统。

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摘要

Correct interactions with extracellular matrix are essential to human pluripotent stem cells (hPSC) to maintain their pluripotent self-renewal capacity during in vitro culture. hPSCs secrete laminin 511/521, one of the most important functional basement membrane components, and they can be maintained on human laminin 511 and 521 in defined culture conditions. However, large-scale production of purified or recombinant laminin 511 and 521 is difficult and expensive. Here we have tested whether a commonly available human choriocarcinoma cell line, JAR, which produces high quantities of laminins, supports the growth of undifferentiated hPSCs. We were able to maintain several human pluripotent stem cell lines on decellularized matrix produced by JAR cells using a defined culture medium. The JAR matrix also supported targeted differentiation of the cells into neuronal and hepatic directions. Importantly, we were able to derive new human induced pluripotent stem cell (hiPSC) lines on JAR matrix and show that adhesion of the early hiPSC colonies to JAR matrix is more efficient than to matrigel. In summary, JAR matrix provides a cost-effective and easy-to-prepare alternative for human pluripotent stem cell culture and differentiation. In addition, this matrix is ideal for the efficient generation of new hiPSC lines.
机译:与细胞外基质的正确相互作用对于人类多能干细胞(hPSC)在体外培养期间维持其多能自我更新能力至关重要。 hPSC分泌层粘连蛋白511/521,这是最重要的功能性基底膜成分之一,它们可以在规定的培养条件下维持在人层粘连蛋白511和521上。但是,纯化或重组层粘连蛋白511和521的大规模生产是困难且昂贵的。在这里,我们测试了产生大量层粘连蛋白的常见人类绒癌组织JAR是否支持未分化hPSC的生长。我们能够使用定义的培养基在JAR细胞产生的脱细胞基质上维持几种人类多能干细胞系。 JAR矩阵还支持将细胞定向分化为神经元和肝细胞方向。重要的是,我们能够在JAR基质上衍生出新的人诱导多能干细胞(hiPSC)品系,并表明早期hiPSC菌落对JAR基质的粘附比对基质胶的粘附更为有效。总而言之,JAR基质为人类多能干细胞培养和分化提供了一种经济高效且易于制备的替代品。此外,该矩阵对于高效生成新的hiPSC线非常理想。

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