High-efficiency non-integrating human iPSC induction platform

Abstract

The invention discloses a highly efficient non-integrated human iPSC induction platform. Composition comprising a pre-inducer and a post-inducer for inducing human cells into iPSCs, the active ingredients of the pre-inducer being: transforming growth factor beta inhibitor, glycogen synthase kinase 3 inhibitor, cAMP agonist , S-adenosyl homocysteine ​​hydrolase inhibitor and p21-activating kinase inhibitor; the active ingredients of the post-inducer are: glycogen synthase kinase 3 inhibitor, selective ATP non-competitive MEK inhibitor and S ‑Adenosyl Homocysteine ​​Hydrolase Inhibitors. Under the action of the pro-inducer composition, the pre-induction efficiency of iPSCs reached 6.4%. By post-induction culture, the whole culture can be induced up to 20.8%, or all pre-inducer clones can be further induced into ESC-like human iPSC clones. The induction efficiency is much higher than that of the existing technology, and the obtained iPSCs have high maturity, no exogenous gene insertion, and the cell shape and performance are closer to ESCs, with good stability and high application value.

Description

technical field [0001] The invention relates to an inducer, an induction method and an induction medium for inducing human somatic cells into iPSCs. Background technique [0002] The successful induction of iPSCs is a major breakthrough in stem cell research. iPSCs have solved the ethical issues in human ESC research, avoided the problem of lack of human oocytes in nuclear transfer technology, and provided new clinical applications for stem cells and regenerative medicine. At the same time, it also provides an important research platform for developmental biology, disease occurrence and development mechanism, gene and protein function research, and drug screening. [0003] There are many methods for inducing somatic cells into iPSCs, such as gene knockout, integration of exogenous genes, cytokine induction or their combination. Gene knockout and integration of exogenous genes in the genome will change the genome, resulting in unstable iPSCs and a great possibility of cancer...

AI Technical Summary

Problems solved by technology

[0006] Due to the large differences in genetic background and culture conditions, our and other studies have shown that the iPSC induction conditions suitable for mice cannot be applied to the induction of human iPSCs, and there is no successful report of chemical induction of human iPS cells

The currently developed induction methods such as free body vectors can also be successfully used to induce human iPSCs and are relatively safe, but the induction efficiency is lower than that of mouse iPS cells, only 0.1-1%.

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