Application of melatonin in preparation of medicine for preventing premature senility of mesenchymal stem cells

A technology of stem cells and melatonin, applied in the field of medical applications, can solve problems such as the impact that has not yet been elucidated, and achieve the effects of increasing cell proliferation potential, enhancing osteogenic differentiation ability, and reducing the positive rate of staining

Inactive Publication Date: 2015-09-02
THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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Problems solved by technology

[0006] Technical issues solved: Aiming at the defect that the effect of melatonin on premature aging mesenchymal stem cells caused by oxidative stress has not yet been elucidated, the present invention provides an application of melatonin in the preparation of a drug for preventing premature aging of mesenchymal stem cells

Method used

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  • Application of melatonin in preparation of medicine for preventing premature senility of mesenchymal stem cells
  • Application of melatonin in preparation of medicine for preventing premature senility of mesenchymal stem cells
  • Application of melatonin in preparation of medicine for preventing premature senility of mesenchymal stem cells

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Embodiment 1

[0032] 1. The effect of melatonin on the proliferation ability, positive rate of SA-β-Gal staining and cell cycle of premature aging mesenchymal stem cells.

[0033] Human BM-MSCs were stored at 37°C, 5% CO 2 Cultivated at 175cm in the incubator 2 In the culture bottle (the volume ratio of each component of the medium is 89% α-MEM, 10% fetal bovine serum, 1% penicillin, streptomycin), the medium is changed every three days. When the cell density is ≥90%, digest the cells with 0.25% trypsin-EDTA at 3000 / cm 2 They were inoculated in well plates of three sizes (96-well plates for proliferation experiments, 12-well plates for SA-β-gal staining, and 6-well plates for cell cycle detection) for the next experiment. When the cell density reaches 50%, add 200 μM HO 2 o 2 (diluted in growth medium), incubate for 2 h. After 2 hours, wash twice with serum-free α-MEM medium, and then add melatonin with different concentration gradients (10nM, 1uM, 100uM) in complete medium to contin...

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Abstract

The invention provides application of melatonin in preparation of a medicine for preventing premature senility of mesenchymal stem cells; the mesenchymal stem cells are mesenchymal stem cells obtained from human, pig, mouse or rabbit tissues; influence of the melatonin to the multiplication capacity of premature senility mesenchymal stem cells is detected by using CCK-8; change of the SA-beta-Gal dyeing positive rate after the melatonin acts on the premature senility mesenchymal stem cells is detected and analyzed by using SA-beta-Gal dyeing; change of the cell cycle after the melatonin acts on the premature senility mesenchymal stem cells is detected and analyzed by using a flow cytometry; dyeing is carried out by using calcium nodule alizarin red; and influence of the melatonin to the osteogenesis ability of the premature senility mesenchymal stem cells is quantitatively detected and analyzed by using calcium nodule. Application disclosed by the invention provides a policy for preventing oxidative stress injury through cell therapy based on the mesenchymal stem cells.

Description

technical field [0001] The invention belongs to the field of medical applications, in particular to the application of melatonin in the preparation of medicines for preventing premature aging of mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) were originally discovered in the bone marrow stroma. Due to their strong self-renewal and multi-lineage differentiation potential, they have been widely used in the fields of tissue engineering and regenerative medicine. In order to achieve therapeutic purposes, MSCs must be expanded in vitro to obtain sufficient numbers, but MSCs have to face the fate of replicative senescence when expanded in vitro. [0003] Cell senescence is a phenomenon in which cells gradually lose their ability to proliferate and the cell cycle stagnates in the G1 phase. In addition, it has been reported that cells subjected to oxidative stress may undergo a premature aging process. Progeria cells exhibit the same functional...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4045A61P39/06A61P39/00
Inventor 何帆周龙刘韬陈曦张文罗宗平杨惠林
Owner THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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