Feeder-free and serum-free human induced pluripotent stem cell isolation culture method

Abstract

The invention relates to a feeder-free and serum-free human induced pluripotent stem cell isolation culture method. By the feeder-free and serum-free human induced pluripotent stem cell isolation culture method, feeder cells and serum are not needed, a culture medium has clear components, operation is simple, human iPS cells can be efficiently and stably isolated, self-renewal and pluripotency are maintained, and a good experimental foundation is laid for final establishment of a clinical application-level human iPS cell isolation culture system.

Description

technical field [0001] The invention relates to the field of cell engineering, in particular to a feeding layer-free and serum-free method for separating and culturing human induced pluripotent stem cells. Background technique [0002] Nowadays, in the commonly used induced pluripotent stem cells (iPScells) isolation and culture system, it largely depends on feeder cells and serum, which are responsible for the self-renewal and pluripotency of iPS cells. plays an important role (Zhou et al., 2015). [0003] The application of feeder cells in pluripotent cell culture has a long history. As early as in vitro culture of teratoma cells (embryonic carcinoma cells, ECC), the use of mouse embryonic fibroblasts (MEFs) that lost mitotic ability ) as feeder cells, and has been used in mouse and human embryonic stem cells (Zwaka and Thomson, 2005). Currently commonly used mouse feeder cells include two types: MEFs and STO cells. Feeder cells may play two roles in maintaining the sel...

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Problems solved by technology

[0006] It is precisely because the feeder layer cells and serum are from other animals, their composition is very complex, the composition is unknown, and the quality is unstable, so the current iPS cell culture system not only has potential safety hazards, it is difficult to cultivate iPScells for clinical application on a large scale, but also for The interpretation of the experimental results also brings a lot of trouble

[0007] In conclusion, the isolation and culture system of human induced pluripotent stem cells (iPS cells) largely depends on feeder cells and serum. This isolation and culture system has potential safety hazards, and it is difficult to cultivate iPS cells on a large scale for clinical application.

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