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Frozen stock solution used for preserving adult stem cells for long time and preparation method thereof

A technology for long-term preservation of adult stem cells, which is applied in the field of cryopreservation for long-term preservation of adult stem cells and the preparation thereof, and can solve the problems of single protective components and the like

Active Publication Date: 2014-04-09
北京清美联创干细胞科技有限公司
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Problems solved by technology

[0004] In order to solve the problem of the single protective component in the existing cell cryopreservation solution, the present invention uses total flavonoids of Epimedium (Xi’an Grass Root Bioengineering Co., Ltd., 20100806) as an additive, and replaces conventional fetal bovine with Knock-out serum (Gibco, 10828028) Serum, improve the anti-apoptosis and anti-oxidation ability of adult stem cells after resuscitation, and remove the influence of exogenous animal-derived serum on cells

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  • Frozen stock solution used for preserving adult stem cells for long time and preparation method thereof

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Embodiment Construction

[0010] Preparation of basic freezing solution:

[0011] Preparation sequence of freezing solution: first add dimethyl sulfoxide to DMEM medium, and then add Knock-out serum;

[0012] Preparation ratio: DMEM medium, dimethyl sulfoxide, and serum are prepared in a ratio of 7:1:2 to prepare the basic cryopreservation solution;

[0013] Preparation of Epimedium total flavonoids frozen liquid:

[0014] Add 1mg of total flavonoids of Epimedium to 10ml of basic freezing solution to prepare a mother solution of 105ng / ml;

[0015] Then take 1 μl of the mother solution and dilute it into 10 ml of the above-mentioned basic freezing solution to obtain the freezing solution with a final concentration of 10 ng / ml.

[0016] Cell Freezing:

[0017] The cells were centrifuged at 1000rpm for 10min to discard the supernatant, resuspended in the cryopreservation solution containing total flavonoids of Epimedium, and put into a sterile cell cryopreservation tube.

[0018] Freezing procedure: ...

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Abstract

The invention relates to a frozen stock solution used for preserving adult stem cells for a long time, and a preparation method thereof. Every 100ml of base frozen stock solution contains: 10-20ng / ml of herba epimedii total flavones, and every 100ml of base frozen stock solution contains: 50-70% of DMEM (dulbecc' modified eagle medium) cluture medium, 10% of dimethyl sulfoxide, and 40-20% of Knock-out serum. The frozen stock solution can improve capacities for preventing cells from frozen damage and resisting oxidation and apoptosis after the cells are recovered, prevents the cells from contacting with heterologous animal original serum during a frozen stock process and further increases security of the adult stem cells for clinic applications.

Description

technical field [0001] The invention belongs to the field of cell tissue engineering, and relates to a cryopreservation solution for long-term preservation of adult stem cells and a preparation method thereof. Background technique [0002] Epimedium total flavonoids (EF) is an extract of traditional Chinese medicine, which can promote the proliferation, migration and antioxidant capacity of stem cells. Using gene chip technology, it was found that EF has a significant effect on corticosterone rat growth hormone (GH), growth hormone releasing hormone (GHRH) and various growth-promoting factors such as insulin-like growth factor binding protein (IGFBP) and nerve growth factor (NGF). Upregulation. In natural aging rats (kidney deficiency syndrome model), it was found that EF can rejuvenate the expression of genes in multiple tissues, and also up-regulate the expressions of GH, GHRH, IGFBP, and NGF, which are down-regulated in aged rats. The embryonic mouse neural stem cells i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
Inventor 田杰
Owner 北京清美联创干细胞科技有限公司
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