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Reprogramming medium for human induced pluripotent stem cell

A technology of pluripotent stem cells and culture medium, applied in the field of reprogramming medium of human induced pluripotent stem cells, can solve the problems of abnormal karyotype, cloned gene rearrangement, troublesome operation, etc., and achieve the effect of improving speed and efficiency

Inactive Publication Date: 2018-07-10
安徽中盛溯源生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Since the virus / plasmid vectors used to transduce foreign genes into donor cells will affect them in the reprogramming process, there are gene rearrangements, karyotype abnormalities, and epigenetic abnormalities in the clones screened by this method. Abnormalities and other phenomena, even with high risk of cancer
In 2008, Okita et al. obtained mouse iPSCs through multiple conventional plasmid transfection methods, but the operation was cumbersome and the reprogramming efficiency was low
[0026] However, hiPSC reprogramming is inefficient in trophoblast-free, serum-free, KSR-free iPSC media such as mTeSR1

Method used

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  • Reprogramming medium for human induced pluripotent stem cell
  • Reprogramming medium for human induced pluripotent stem cell
  • Reprogramming medium for human induced pluripotent stem cell

Examples

Experimental program
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Effect test

Embodiment 1

[0059] Example 1. Reprogramming of human erythrocyte progenitor-derived hiPSCs

[0060] 1. Preparation of reprogramming medium:

[0061] The formula of the reprogramming medium provided by the present invention is shown in Table 1. The erythrocyte progenitor cell medium is taken, and other components are weighed, dissolved, filtered, and added to the medium one by one. Erythroid progenitor medium without any small molecules was used as a control medium.

[0062] Table 1 Reprogramming medium formulations with different ratios

[0063]

[0064] Among them, P represents PD0325901; C represents CHIR99021; A represents A-83-01; PC represents the combination of PD0325901 and CHIR99021; CA represents the combination of CHIR99021 and A-83-01; PCA represents the combination of PD0325901, CHIR99021 and A-83-01.

[0065] 2. To detect the reprogramming efficiency of erythrocyte group cells induced by the medium to hiPSC, the following methods were used:

[0066] a. Reprogramming pla...

Embodiment 2

[0071] Example 2. Reprogramming of human fibroblast-derived hiPSCs

[0072] 1. Preparation of reprogramming medium:

[0073] The formula of the reprogramming medium provided by the present invention is shown in Table 2. Take the LONZA MSC serum-free medium, weigh the other components, dissolve, filter, and add to the medium one by one, mix well and store at 4°C. In addition, LONZA MSC serum-free medium without any small molecules was used as a control medium.

[0074] Table 2 Reprogramming medium formulations with different ratios

[0075]

[0076] 2. To detect the reprogramming efficiency of culture medium for inducing human fibroblasts into hiPSCs, the following methods can be used:

[0077] a. Reprogramming plasmid transfection: After the human fibroblasts are full, take 0.5-2×10 cells 6 , a total of 4 groups, of which 3 groups are experimental groups and 1 group is a control group. The plasmids carrying the reprogramming factors were electrotransfected into the cells...

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Abstract

The invention belongs to the technical field of culture media, and particularly relates to a reprogramming medium for a human induced pluripotent stem cell (hiPSC). The reprogramming medium comprisesan adult cell culture medium for culturing a human somatic cell and a cell signaling pathway inhibitor added to the adult cell culture medium; and the cell signaling pathway inhibitor is any one of ora combination containing more of a glycogen synthase kinase 3 inhibitor, a mitogen-activated protein kinase kinase inhibitor and a transforming growth factor beta inhibitor. The reprogramming mediumis obtained through adding one or more small-molecular compounds to adult cell culture medium, is used for induced reprogramming culture or induced differentiation culture, is a non-trophoblastic animal origin-free medium, greatly improves the speed and the efficiency of reprogramming of the human somatic cell into the human pluripotent stem cell, and especially can efficiently reprogram the humansomatic cell into the hiPSC for expressing specific markers.

Description

technical field [0001] The invention belongs to the technical field of medium, and in particular relates to a reprogramming medium for human induced pluripotent stem cells. Background technique [0002] Stem cells refer to a type of cell that is capable of self-renewal, division, and differentiation into other cell types. Judging from the stage of stem cell development, stem cells can differentiate into embryonic stem cells (embryonicstem cells, ESCs) and adult stem cells (adult stem cells). Embryonic stem cells are a kind of pluripotent stem cells (pluripotent stem cells), which can proliferate indefinitely and differentiate into more than 200 types of cells in the human body, forming tissues and organs of the body, that is, stem cells with the potential to differentiate into various cells and tissues. Adult stem cells include those present in adult tissues such as bone marrow mesenchymal stem cells, blood stem cells, and neural stem cells. [0003] In 2006, the Yamanaka ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074
CPCC12N5/0696C12N2501/15C12N2501/727
Inventor 俞君英董成友张颖
Owner 安徽中盛溯源生物科技有限公司
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