High-efficiency non-integrated human iPSC induction platform

A technology of inducer and induction medium, applied in the field of high-efficiency non-integrated human iPSC induction platform, which can solve the problems of no successful report of chemical induction of human iPS cells, large differences in genetic background and culture conditions, and inability to apply human iPSC induction , to achieve the effects of high induction efficiency, high maturity and good stability

Active Publication Date: 2015-06-03
GUANGDONG HOSPITAL OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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. Successful reports of chemical

Method used

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  • High-efficiency non-integrated human iPSC induction platform
  • High-efficiency non-integrated human iPSC induction platform
  • High-efficiency non-integrated human iPSC induction platform

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Embodiment Construction

[0036] A composition for inducing human cells into iPSCs, consisting of a pre-inducer and a post-inducer, wherein,

[0037] The active ingredients of the pro-inducer are: transforming growth factor beta inhibitors, glycogen synthase kinase 3 inhibitors, cAMP agonists, S-adenosyl homocysteine ​​hydrolase inhibitors and p21-activated kinase inhibitors;

[0038] The active ingredients of the post-inducer are: glycogen synthase kinase 3 inhibitor, selective ATP non-competitive MEK inhibitor and S-adenosyl homocysteine ​​hydrolase inhibitor.

[0039] As a further improvement of the present invention, in the above pre-inducer and post-inducer, the transforming growth factor beta inhibitor is selected from E-616452, SB431542, LDN193189, Dorsomorphin, LY2109761, LY2157299, SB525334, SB505124, GW788388, LY364947, D4476, IDE1 , ITD1, SD208, A83-01.

[0040] As a further improvement of the present invention, in the above composition, the glycogen synthase kinase 3 inhibitor is selected ...

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Abstract

The invention discloses a high-efficiency non-integrated human iPSC induction platform which comprises a composition, wherein the composition is used for inducing a human cell into iPSC, and the composition comprises a previous inducer and a later inducer; the previous inducer comprises the following active components: a transforming growth factor beta inhibitor, a glycogen synthetase kinase 3 inhibitor, a cAMP agonist, an S-adenosyl homocysteine hydrolase inhibitor and a p21 activated kinase inhibitor; and the later inducer comprises the following active components: the glycogen synthetase kinase 3 inhibitor, a selective ATP noncompetitive MEK inhibitor and the S-adenosyl homocysteine hydrolase inhibitor. The high-efficiency non-integrated human iPSC induction platform disclosed by the invention achieves the previous induction efficiency of the iPSC of 6.4% under the action of a previous inducer composition, can achieve the induction of a full culture of 20.8% through later induction culture or be used for totally further inducing a previous inductor clone into a human iPSC clone approaching to ESC and is far higher than the prior art in induction efficiency; in addition, the obtained iPSC is high in maturity, free of being inserted with an exogenous gene, is more approaching to the ESC in cell morphology and property and has the advantages of good stability and great 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...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N15/85
Inventor 林同香徐洋吴寿海
Owner GUANGDONG HOSPITAL OF TRADITIONAL CHINESE MEDICINE
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