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Reprogramming culture medium, preparation method thereof and culture method of reprogramming cell

A reprogramming and culture medium technology, applied in the field of biomedicine, can solve problems such as prone to human rejection and affect the efficiency of cell reprogramming, and achieve the effect of stable quality, high reprogramming efficiency, and clear components

Active Publication Date: 2014-02-26
BEIJING CELLAPY BIOTECH
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Problems solved by technology

[0004] The purpose of the present invention is to provide a reprogramming medium, its preparation method, and a method for culturing reprogrammed cells, so as to solve the problem that hiPS cells in the prior art are prone to produce human body after transplantation due to long-term exposure to a culture environment rich in animal products. Rejection reaction and the technical problem that the reprogramming medium in the prior art seriously affects the efficiency of cell reprogramming

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  • Reprogramming culture medium, preparation method thereof and culture method of reprogramming cell
  • Reprogramming culture medium, preparation method thereof and culture method of reprogramming cell
  • Reprogramming culture medium, preparation method thereof and culture method of reprogramming cell

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Embodiment approach

[0026]Preferably, the reprogramming medium is prepared into reprogramming medium 1 and reprogramming medium 2 before use, and the configuration ratio can be adjusted according to the actual situation. According to a typical implementation of the present invention, the following steps are specifically included: Thaw reprogramming supplement 1 and reprogramming supplement 2 at 2-8°C; mix reprogramming supplement 1 and basal medium at a volume of 7:500 to form reprogramming medium 1; mix reprogramming supplement 2 and basal medium Reprogramming medium 2 was formed after mixing at a volume of 7:500. Preferably, reprogramming medium 1 and reprogramming medium 2 are prepared 0-14 days before use and stored at 2-8°C.

[0027] According to a typical embodiment of the present invention, a method for culturing reprogrammed cells is provided. The culture method is to use the above-mentioned reprogramming medium for culture.

[0028] Preferably, the following steps are included: S1, see...

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Abstract

The invention discloses a reprogramming culture medium, a preparation method thereof and a culture method of a reprogramming cell, wherein the reprogramming culture medium comprises a basal culture medium, a reprogramming additive 1 and a reprogramming additive 2, wherein the basal culture medium is a DMEM (Dulbecco Modified Eagle Medium) / F12 culture medium; the reprogramming additive 1 comprises sodium hydrogen carbonate, sodium selenate, recombinant human insulin, recombinant human basic fibroblast growth factors, recombinant human transferrin, ascorbic acid and hydrocortisone; the reprogramming additive 2 comprises sodium hydrogen carbonate, sodium selenate, recombinant human insulin, recombinant human basic fibroblast growth factors, recombinant human transferrin, ascorbic acid, valproic acid and sodium butyrate. The reprogramming culture medium disclosed by the invention is exact in component, stable in quality, high in reprogramming efficiency, wide in cell lineage and capable of reprogramming and culturing cells such as human skin fibroblasts, peripheral blood mononuclear cells, amniotic epithelial cells and the like.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to a reprogramming medium, a preparation method thereof and a method for culturing reprogrammed cells. Background technique [0002] In 2006 and 2007, Japanese scholar Shinya Yamanaka successfully introduced four transcription factors (Oct4, Sox2, Klf4 and c-Myc) into mice respectively (Takahashi and Yamanaka., 2006) and human (Takahashi et al., 2007) skin fibroblasts were reprogrammed into induced pluripotent stem cells (Induced Pluripotent Stem Cells, iPSCs), and thus won the 2012 Nobel Prize in Medicine and Physiology. hiPS cells (induced pluripotent stem cells) have all the differentiation capabilities of hES cells (human embryonic stem cells) and have no ethical issues. They will completely replace hES cells in the near future and become the most important cell source for regenerative medicine. [0003] At present, the reprogramming culture conditions used by most research...

Claims

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Application Information

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IPC IPC(8): C12N5/10
Inventor 李扬兰峰
Owner BEIJING CELLAPY BIOTECH
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