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Application of trichostatin A in maintaining dryness of stem cells

A technology of trichostatin and stem cells, applied in the field of stem cell culture, can solve the problems of reducing and restricting the clinical application of stem cells, unable to maintain stemness, etc., and achieve the effects of good stemness, less autonomous differentiation and slow aging speed of stem cells

Active Publication Date: 2013-11-06
国健清科生物医药科技(北京)有限责任公司
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, stem cells will undergo autonomous differentiation and aging during in vitro expansion, that is, they cannot maintain stemness, thereby reducing their ability to differentiate into other more important cells such as nerve cells and cardiomyocytes, which is a prominent problem in stem cell culture at present , will also seriously restrict the clinical application of stem cells

Method used

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  • Application of trichostatin A in maintaining dryness of stem cells
  • Application of trichostatin A in maintaining dryness of stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Follow the steps below to conduct experiments to study the effects of adding different concentrations of TSA on the culture of MSCs and determine the optimal concentration of TSA:

[0028] Different concentrations of TSA ((0, 6.25, 12.5, 25, 50, 100, 200 and 300 nM) were added to conventional MSCs culture medium (low glucose DMEM plus 10% fetal bovine serum, 100 IU penicillin and 100 μg. / ml streptomycin) ), prepared into stem cell medium, and then in 5% CO 2Under the condition of 37 degrees Celsius and 37 degrees Celsius, the medium was used to culture MSCs, and after 3 days of culture, the cells were harvested and counted.

[0029] See the results of cell counts after TSA treatment figure 1 . like figure 1 As shown, where * means Pfigure 1 The results showed that the number of MSCs increased after low concentration (6-50nM) TSA treatment. Thus, it was shown that the addition of 6-50 nM trichostatin A could effectively promote the proliferation of MSCs without produ...

Embodiment 2

[0031] According to the experimental results of Example 1, the best TSA concentration for MSC proliferation was selected for further long-term effect test:

[0032] MSCs were continuously subcultured (from the 1st passage to the 6th passage) with a culture solution containing 6.25 nM TSA (other specific components were the same as in Example 1), and the cell count was performed using a cell counter to calculate the cumulative number of cells. Among them, the culture solution containing DMSO but no TSA was used as the blank control. For the statistical results of the cumulative number of cells in each generation, see figure 2 . like figure 2 Shown, where * means P figure 2 The results showed that at passage 6, the number of MSCs cultured in TSA-containing medium increased 10-fold compared with the number of MSCs cultured in blank control (*P<0.01). Thus, it was proved that the addition of 6.25 nM trichostatin A has a significant effect on inhibiting MSC senescence and main...

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Abstract

The invention discloses application of trichostatin A (TSA) in maintaining dryness of stem cells, a stem cell medium, a kit used for culture of stem cells and a stem cell culture method. According to the invention, a certain dosage of, e.g., 6 to 50 nM, TSA is added in a traditional stem cell medium, so the state of histone acetylation in the process of multiplication culture of the stem cells can be effectively maintained; and thus, dryness of the stem cells can be effectively maintained, and autonomous differentiation of the stem cells during in vitro culture is avoided.

Description

technical field [0001] The invention relates to the technical field of stem cell culture, in particular, the invention relates to the use of trichostatin A in maintaining the stemness of stem cells. More specifically, the present invention relates to the use of trichostatin A in maintaining the stemness of stem cells, a stem cell culture medium, a kit for stem cell culture, and a stem cell culture method. Background technique [0002] Stem cells, especially mesenchymal stem cells (mesenchymal stem cells, or mesenchymal stromal cells, MSCs), can promote the repair of various damaged tissues and have broad clinical application prospects. However, stem cells will undergo autonomous differentiation and aging during in vitro expansion, that is, they cannot maintain stemness, thereby reducing their ability to differentiate into other more important cells such as nerve cells and cardiomyocytes, which is a prominent problem in stem cell culture at present , will also seriously rest...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 吴耀炯李智龙
Owner 国健清科生物医药科技(北京)有限责任公司
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