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Feeder-free and xeno-free culture of human pluripotent stem cells using UCBS matrix

机译：使用UCBS基质对人多能干细胞进行无饲养和无异种培养

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摘要

The ideal medium for human pluripotent stem cells (hPSCs) culture should be feeder-free, xeno-free, and completely defined. The present study aims to establish a new feeder-free and xeno-free system for culturing hPSCs. The system consists of the matrix, which was prepared from human umbilical cord blood serum (UCBS) and used to coat the culture plates, and the xeno-free medium, which was conventional serum-free hES medium supplemented with high concentrations of bFGF and Fibronectin. Compared with matrigel and mouse embryonic fibroblasts (MEFs), the UCBS matrix was proved to be equally suitable for the growth of hPSCs. After a series of experiments with different media and cytokins, the UCBS matrix was found worked the best with the basic medium (BM) supplemented with 20 ng/mL bFGF, 10 ug/mL fibronectin and Y-27632 for culture of hES cells. The hPSCs maintained normal karyotype, high proliferation rate and self-renewal ability after continuous culture more than 10 passages in this feeder-free and xeno-free system. Furthermore, a new human embryonic stem (hES) cell line was derived from discarded day 3 embryos in this newly developed culture system. In conclusion, this feeder-free and xeno-free system could not only be used to the culture hPSCs, but could also be used to derive new hES cell lines.
机译：人类多能干细胞（hPSC）培养的理想培养基应无饲养层，无异种且完整定义。本研究旨在建立新的无饲养者和无异种培养hPSC的系统。该系统由从人脐带血血清（UCBS）制备并用于覆盖培养板的基质和无异种培养基组成，后者是常规的无血清hES培养基，并补充有高浓度的bFGF和纤连蛋白。 。与基质胶和小鼠胚胎成纤维细胞（MEF）相比，UCBS基质同样适用于hPSC的生长。经过一系列不同培养基和细胞因子的实验后，发现UCBS基质在添加20 ng / mL bFGF，10 ug / mL纤连蛋白和Y-27632的基础培养基（BM）的基础上最有效地培养hES细胞。在无饲养者和无异种系统的条件下，连续培养超过10代后，hPSC保持了正常的核型，高增殖率和自我更新能力。此外，在这种新开发的培养系统中，从废弃的第3天胚胎中获得了新的人类胚胎干（hES）细胞系。总之，这种无饲养层和无异种系统不仅可以用于培养hPSC，而且还可以用于衍生新的hES细胞系。

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