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Cell reprogramming method

A reprogramming and cell technology, applied in animal cells, vertebrate cells, genetically modified cells, etc., can solve the problems of non-uniform cloning, low efficiency, and difficulty in automation, and achieve high cloning quality, reduce difficulty, and facilitate tracking. Effect

Inactive Publication Date: 2019-04-23
BEIHANG UNIV
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Problems solved by technology

At present, induced pluripotent stem cell technology has matured, and its clinical and industrial transformation will be the future development direction. However, the current reprogramming method still has low efficiency. See reference [2]: M.Stadtfeld, K.Hochedlinger, "Induced pluripotency: history, mechanisms, and applications," vol.5, pp.582–596, 2010, strong randomness, resulting in uneven clones, see reference [3]: Z.D.Smith, C.Sindhu, and A.Meissner, "Molecular features of cellular reprogramming and development," Nat. Rev. Mol. Cell Biol., vol.17, no.3, pp.139–154, 2016., Difficult to automate and other issues

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Embodiment Construction

[0034] A novel high-efficiency cell reprogramming method provided by the present invention will be described in detail below with reference to the accompanying drawings and examples.

[0035] In the embodiment of the present invention, the limited growth area in the microporous structure increases the occurrence of intercellular adhesion, promotes MET (mesenchymal-epithelial transition), and finally improves the efficiency of iPSCs generation. Finally, the present invention obtains more than 90% of Nanog-positive cell clusters on the superhydrophobic microwell array chip (hereinafter referred to as the chip, which has a micropore structure), and the expression of the MET marker gene is significantly higher than that of the traditional cell culture dish. The reprogramming efficiency was improved by 5 times. Second, cell tracking is more convenient in the microwells of the superhydrophobic microwell array chip than in traditional cell culture dishes. In the experiment, the aggr...

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Abstract

The invention discloses a cell reprogramming method and belongs to the technical field of induced pluripotent stem cells. The method comprises the steps of on-chip cell reprogramming, on-chip clone selection and on-chip microenvironment optimization. The method can obviously improve the cell reprogramming efficiency. Produced clones have higher quality, and the occurrence positions of the clones are controllable and more uniform instead of being random as before, so that realization of subsequent industrialized standards is facilitated. By means of the method, cells can be conveniently tracked, the dynamic change process of reprogramming is monitored, and therefore convenience is provided for understanding the mechanism of reprogramming. The method can achieve precise control over the micro-pore physical and chemical environment at high flux, and screening at the high flux is favorable for the culture micro-environment of reprogramming.

Description

technical field [0001] The invention belongs to the technical field of induced pluripotent stem cells, relates to cell reprogramming, and specifically refers to an efficient cell reprogramming method based on a superhydrophobic micropore array chip. Background technique [0002] Induced pluripotent stem cells (iPSCs: induced pluripotent stem cells) have broad application prospects in regenerative medicine, drug screening, disease models, etc. Since they were first reported in 2006, they have attracted widespread attention from the scientific community. See References [1]: Takahashi K.Yamanaka S.Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors[J].Cell,2006,126:663-676. At present, induced pluripotent stem cell technology has matured, and its clinical and industrial transformation will be the future development direction. However, the current reprogramming method still has low efficiency. See reference [2]: M.Stadtfeld, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12M3/00
CPCC12M23/12C12N5/0656C12N2510/00
Inventor 陈晓芳瞿佳楠
Owner BEIHANG UNIV
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