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Organism lifetime based method for screening compounds capable of improving generation efficiency of induced pluripotent stem cells

A technology for pluripotent stem cells and candidate compounds, which is applied in the field of screening compounds that improve the production efficiency of induced pluripotent stem cells based on the lifespan of organisms, and can solve problems such as low efficiency, affecting the promotion and application of induced pluripotent stem cells, and low efficiency.

Active Publication Date: 2015-02-18
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] From previous research results, the efficiency of induced pluripotent stem cells is basically the same, and the efficiency is about ten induced pluripotent stem cells from one hundred thousand starting cells, which is very low
Low efficiency will seriously affect the promotion and application of induced pluripotent stem cells
Since the generation of induced pluripotent stem cells, the exploration of its reprogramming mechanism and high efficiency has not stopped. Current research has proved that some compounds can improve the production efficiency of induced pluripotent stem cells by regulating chromatin structure. However, in the field of There is still a great need for compounds that increase the efficiency of induced pluripotent stem cell generation, and there is a need for more efficient systems for screening compounds that increase the efficiency of induced pluripotent stem cell generation

Method used

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  • Organism lifetime based method for screening compounds capable of improving generation efficiency of induced pluripotent stem cells
  • Organism lifetime based method for screening compounds capable of improving generation efficiency of induced pluripotent stem cells
  • Organism lifetime based method for screening compounds capable of improving generation efficiency of induced pluripotent stem cells

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Experimental program
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Effect test

Embodiment 1

[0069] Effect of embodiment 1 rapamycin, PP242 and PQ401 on the lifespan of fruit flies

[0070] The inventors tested the effects of 3 μM rapamycin, different concentrations of PP242 and different concentrations of PQ401 on the lifespan of Drosophila according to the above method. It was found that 3 μM rapamycin, 0.3 μM, 1 μM and 3 μM PP242, and 3 μM, 10 μM and 30 μM PQ401 could significantly increase the survival percentage of Drosophila (eg figure 1 C and E) and average survival time (as shown in figure 1 D and F shown).

Embodiment 2

[0071] Embodiment 2 PP242 and PQ401 to mouse fibroblast GFP + percent impact

[0072] The inventors detected the GFP produced by SKOM transfection under the action of different concentrations of PP242 and different concentrations of PQ401 according to the above method + Cell percentage and GFP + The number of colonies was used to determine the effects of PP242 and PQ401 on the successful transfection of SKOM into mouse fibroblasts. The results showed that PP242 at 0.03nM, 0.1nM, 0.3nM and 1nM, and PQ401 at 0.3nM, 1nM, 3nM and 10nM all increased GFP compared to controls not treated with the subject compound + Cell percentage and GFP + The number of colonies increases the efficiency of SKOM to successfully transfect mouse fibroblasts and make them into induced pluripotent stem cells (such as figure 1 A and B shown). Through quantitative analysis, it can be determined that these compounds can increase the induction efficiency of induced pluripotent stem cells by as much as 3...

Embodiment 3

[0073] Embodiment 3 various compounds are to mouse fibroblast GFP + percent impact

[0074] The inventors also selected rapamycin and compounds that have life-prolonging effects on model organisms such as yeast, nematodes, Drosophila, and mice reported in the literature, including resveratrol, fisetin, spermidine, LY294002 , curcumin and metformin [14-18] , GFP produced by transfection with SKOM under their action was detected according to the method described above + cell percentage. It was surprisingly found that these compounds (3 μM resveratrol, 3 μM fisetin, 30 nM spermidine, 0.3 μM LY294002, 10 μM curcumin) were GFP + The percentage of cells, that is, the efficiency of successfully transfecting mouse fibroblasts with SKOM to make them into induced pluripotent stem cells (such as figure 1 G and H).

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Abstract

The invention relates to a method for screening compounds capable of improving generation efficiency of induced pluripotent stem cells, in particular to a method for screening compounds capable of improving generation efficiency of induced pluripotent stem cells in basis of organism lifetimes, particularly fruit fly lifetimes. The invention further relates to a lifetime prolonging compound, in particular to a fruit fly lifetime prolonging compound, and particularly discloses new usages of rapamycin, PP242, PQ401, resveratrol, fisetin, spermidine, LY294002 and curcumin in improvement of generation efficiency of induced pluripotent stem cells.

Description

technical field [0001] The present invention relates to a method for screening compounds that improve the production efficiency of induced pluripotent stem cells (iPSC for short), specifically based on the lifespan of organisms, especially the lifespan of fruit flies to screen for compounds that improve the production efficiency of induced pluripotent stem cells method. The present invention also relates to lifespan-extending compounds, in particular Drosophila lifespan-extending compounds, in particular rapamycin, PP242, PQ401, resveratrol, fisetin, spermidine, LY294002 and curcumin in enhancing inducible pluripotency New uses in stem cell production efficiency. Background technique [0002] Embryonic stem cells (ESC) are a group of cells derived from the inner cell mass of the embryonic blastocyst, which can proliferate indefinitely in vitro while maintaining the ability of multilineage differentiation. More than a decade ago, the successful isolation of human embryonic ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02C12N5/071A01K61/00A01K67/02A01K67/033
CPCY02A40/81
Inventor 裴钢于洁赵简陈涛涛沈立万洪江
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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