Method for promoting induced pluripotent stem cells to induce and differentiate to be nerve cells

A technology for pluripotent stem cells and inducing differentiation, which is applied in the field of promoting the induction and differentiation of human induced pluripotent stem cells into neurons, so as to achieve the effect of accelerating the formation of induced differentiation and improving the growth state of neurons

Active Publication Date: 2016-02-17
THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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However, the role of autophagy in the differentiation of SCA3-in

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  • Method for promoting induced pluripotent stem cells to induce and differentiate to be nerve cells
  • Method for promoting induced pluripotent stem cells to induce and differentiate to be nerve cells

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Embodiment

[0036] A method for promoting human induced pluripotent stem cells to differentiate into neurons, comprising the following steps:

[0037] (1) Human induced pluripotent stem cells (iPS) obtained by culturing under trophoblast-free conditions, when the degree of confluence is 70-80%, add preheated After the cell digestion solution, incubate at 37°C for 5 minutes, then pipette several times to disperse the cells into a single cell suspension, and then divide the cells into the culture wells; the division density is 2.5× for each culture well. 10 5 –3.0×10 5 When dividing, 10 μM ROCK inhibitor Y27632 can be added to the culture well to prevent cell death;

[0038] (2) On the first day after sorting, aspirate the supernatant of the culture well to remove unattached cells, add preheated Neural induction complete medium, placed in an incubator to continue culturing; thereafter, the medium was changed every other day, P0 generation neural stem cells (NSCs) could be obtained on t...

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Abstract

The invention discloses a method for promoting induced pluripotent stem cells to induce and differentiate to be nerve cells, which comprises steps: induced pluripotent stem (iPS) cells are blown and absorbed to disperse to be single cell suspension after being digested, and are separately planted in culture holes, and rock (ROCK) inhibitors are added into each culture hole; a supernate is adsorbed to discard in the first day after the iPS cells are separately planted, is added into a neural induction complete culture medium, and rapamycin needs to be added into the neural induction complete culture medium to promote autophagy; the neural induction complete culture medium is changed to add low concentration rapamycin to promote autophagy neural cells to differentiate the neural induction complete culture medium after two days, and can be induced to differentiate to be neural cells after being cultured for 12-15 days. The method for promoting the induced pluripotent stem cells to induce and differentiate to be the nerve cells can speed up neurons to induce and differentiate, and improves neuron growth states ofspinocerebellum ataxic three-type iPS.

Description

technical field [0001] The invention relates to a method for promoting the induced differentiation of human induced pluripotent stem cells into neurons, and the method improves the efficiency of induced differentiation by enhancing autophagy. Background technique [0002] Neurons are non-renewable cells, so in some human diseases that damage neurons, some irreversible consequences will occur. With the prolongation of human lifespan, diseases involving neurons such as Alzheimer's disease and cerebrovascular accidents increase, and human induced pluripotent stem cells can be induced to differentiate into functional neurons. possibility. However, induction of differentiation from induced pluripotent stem cells or embryonic stem cells into neurons requires complicated procedures, such as methods involving suspension culture or co-culture of exogenous cells. These methods have the disadvantages of cumbersome operation and long time consumption. However, Drury-Stewart et al. us...

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Application Information

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IPC IPC(8): C12N5/0793C12N5/0797
Inventor 孙筱放欧展辉牛晓华何文茵罗敏陈玉嫦
Owner THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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