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Adipose-derived mesenchymal stem cell transport protection solution and application thereof

A kind of stem cell and protective solution technology, applied in the biological field, can solve the problems of not being able to maintain cell stability and cell activity, and achieve the effects of easy storage and long-distance transportation, safe use, and high cell viability

Inactive Publication Date: 2018-05-22
重庆斯德姆生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a transport protection solution for adipose-derived mesenchymal stem cells, which can solve the shortcomings of existing adipose-derived mesenchymal stem cell transport protection solutions that cannot maintain cell stability and cell activity during long-distance transport.

Method used

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  • Adipose-derived mesenchymal stem cell transport protection solution and application thereof

Examples

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Embodiment 1

[0024] Example 1 Adipose-derived mesenchymal stem cell transport protection solution according to the present invention

[0025] (1) Formula

[0026] The adipose-derived mesenchymal stem cell transport protection solution consists of 10% to 30% DMSO (dimethyl sulfoxide), 50 μg / mL to 150 μg / mL streptomycin, 50% to 80% DMEM (containing various amino acids and glucose) medium), 8% to 20% HES (hydroxyethyl starch).

[0027] (2) Preparation method

[0028] Dissolve streptomycin with DMEM and HES according to the ratio in the formula and mix evenly, then add DMSO, and then place in an ice-water bath until the temperature of the freezing liquid is close to 0°C.

Embodiment 2

[0029] Example 2 Preparation method of human adipose-derived mesenchymal stem cells

[0030] The preparation method of human adipose-derived mesenchymal stem cells comprises the following steps:

[0031] (1) Collect 5-10 g of massive adipose tissue under sterile conditions and soak in DMEM containing antibiotics. Remove the vascular tissue visible to the naked eye, rinse it with DMEM containing antibiotics three times to remove blood cells and other impurities, and cut the tissue block as much as possible with sterile scissors.

[0032] (2) Place adipose tissue in double volume of 0.1% (mass concentration) type I collagenase, shake and digest in a constant temperature water bath at 37°C for 60 minutes, centrifuge at 1500r / min for 10 minutes, discard the supernatant and upper tissue, and add complete culture medium (containing 12% HES (hydroxyethyl starch), 100 μg / mL streptomycin, 70% DMEM) the cells were pelleted into a single-cell suspension by pipetting, filtered through a ...

Embodiment 3

[0034] Example 3 Mixing mode of adipose-derived mesenchymal stem cells and protective solution

[0035] The adipose-derived mesenchymal stem cells are adjusted and added to the adipose-derived mesenchymal stem cell transport protection solution in the form of resuspension, and the concentration of human adipose-derived mesenchymal stem cells in the transport protection solution is 6×10 5 ~7×10 5 pieces / ml. Specifically, the following steps are included: take the third-generation adipose-derived mesenchymal stem cells (ADMSCs) in the logarithmic growth phase, centrifuge at 1000r / min for 5min, discard the supernatant, add the transport protection solution pre-frozen at 4°C, blow and beat the cell pellet, Make the cells evenly distributed in the protection solution, adjust the concentration of viable cells to 6×10 5 ~7×10 5 pc / ml, aliquoted into cryovials for storage.

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Abstract

The invention relates to the technical field of biology, provides a stem cell transport protection solution and particularly relates to an adipose-derived mesenchymal stem cell transport protection solution suitable for long-distance transport. The stem cell transport protection solution is prepared from 10 to 30 percent DMSO (dimethyl sulfoxide), 50 to 150 mu g / mL streptomycin, a 50 to 80 percentDMEM (Dulbecco modified eagle medium) and 8 to 20 percent HES (hydroxyethyl starch). The adipose-derived mesenchymal stem cell transport protection solution has the advantages that the stability of cells and the activity of the cells may be maintained, and convenience is brought to storage and long-distance transport.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a stem cell transport protection solution, in particular to an adipose-derived mesenchymal stem cell transport protection solution suitable for long-distance transport. Background technique [0002] Adipose-derived Mesenchymal Stem Cells (ADSCs) are stem cells derived from adipose tissue. Like other adult stem cells, they have the ability of self-renewal and multidirectional differentiation. Under appropriate induction conditions, they can differentiate into osteoblasts, Fat cells, chondrocytes, etc. are of great value for regenerative medicine and cell therapy research. [0003] Bone marrow mesenchymal stem cells (BMSCs) have become a recognized source of bone seed cells due to their osteogenic ability. However, the application of bone marrow mesenchymal stem cells is often limited due to reasons such as difficult extraction, limited amount of bone marrow extraction, and low extractio...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/0775
CPCA01N1/0221A01N1/0226C12N5/0667
Inventor 胡小东熊华强孙静秦连荣
Owner 重庆斯德姆生物技术有限公司
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