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ADSC (adipose tissue-derived stromal cell) preserving liquid and preparation method and application thereof

A technology of stem cells and preservation solution, applied in the field of stem cells, can solve the problem that it is difficult to ensure that it can be used immediately

Inactive Publication Date: 2019-02-22
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Clinically, the donors and recipients of cell transplantation usually have different timing and location, and it is difficult to ensure that they can be used immediately, and a sufficient number of mesenchymal stem cells is one of the important factors that determine whether clinical efficacy can be achieved.

Method used

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  • ADSC (adipose tissue-derived stromal cell) preserving liquid and preparation method and application thereof
  • ADSC (adipose tissue-derived stromal cell) preserving liquid and preparation method and application thereof

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preparation example Construction

[0020] The present invention also provides a method for preparing adipose-derived mesenchymal stem cell preservation solution, comprising: slowly adding DMSO into fetal bovine serum to obtain adipose-derived mesenchymal stem cell preservation solution. The present invention has no special requirements on the mixing method, and those skilled in the art can choose a suitable mixing method according to common knowledge. Preferably, in the present invention, DMSO is slowly added to the mixture of fetal bovine serum and Lonza DMEM medium to obtain adipose-derived mesenchymal stem cell preservation solution.

[0021] The present invention also provides an application of the adipose-derived mesenchymal stem cell preservation solution described in the present invention as a preservation solution for preparing adipose-derived mesenchymal stem cells.

[0022] The invention provides a fat-derived mesenchymal stem cell preservation solution and its preparation method and application. The ...

Embodiment 1

[0025] All kinds of cryopreservation solutions are sterile-grade products, which are prepared and used immediately. The major components are added first, and DMSO is added slowly to avoid local overheating. Store at 4°C for later use.

[0026] The specific formula is shown in Table 1 to Table 3:

[0027] Table 1, formula one

[0028] DMSO

human serum albumin

DMEM / F12

10wt%

10wt%

80wt%

[0029] Table 2, formula two

[0030] DMSO

FBS (Fetal Bovine Serum)

20wt%

80wt%

[0031] Table 3, formula three

[0032] DMSO

Lonza DMEM medium

FBS

10wt%

20wt%

70wt%

[0033] Add the adipose-derived mesenchymal stem cells to the prepared cryopreservation solution, cool down to -80°C at 1°C / min, and then transfer to liquid nitrogen for storage at -196°C. Then the cryopreserved adipose-derived mesenchymal stem cells were resuscitated and cultured, and the specific resuscitation and culture ste...

Embodiment 2

[0042] Prepare cryopreservation solution according to the formula in Table 5, add adipose-derived mesenchymal stem cells to the prepared cryopreservation solution, cool down to -80°C at 1°C / min, and then transfer to liquid nitrogen for storage at -196°C. Then the cryopreserved adipose-derived mesenchymal stem cells were resuscitated and cultured, and the specific steps of the resuscitated culture were as follows:

[0043] 1) Use a pipette to absorb complete medium solution 10 times the volume of the freezing solution and add it to a 50mL centrifuge tube, quickly transfer the cell suspension to a 50mL centrifuge tube, gently blow and beat to mix, and centrifuge at 500g for 5min.

[0044] 2) Resuspend the cell pellet with complete medium, and adjust the cell seeding density to (5-8)x10 4 cell / mL, add 1μg / mL EGF to a final concentration of 10ng / mL, mix well and inoculate.

[0045] 3) Use a pipette to inoculate 10 mL of cell suspension into a 10 cm culture dish, transfer to 5% CO...

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Abstract

The invention provides an ADSC (adipose tissue-derived stromal cell) preserving liquid and a preparation method and application thereof. The ADSC preserving liquid provided by the invention comprisesfetal calf serum and DMSO; in comparison with the existing preserving liquid, a result shows that the cell viability and corresponding biology characteristics are well protected after the cryopreservation and resuscitation culture of ADSC by taking the preserving liquid provided by the invention used as the ADSC preserving liquid.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a fat-derived mesenchymal stem cell preservation solution and a preparation method and application thereof. Background technique [0002] Adult stem cells are derived from adult tissues, avoiding ethical issues, and are also a kind of cells capable of self-renewal and multi-directional differentiation, so they have broader research and application prospects. Adipose-derived mesenchymal stem cells are stem cells that exist in adipose tissue and have the potential for multilineage differentiation. In 2001, adipose-derived mesenchymal stem cells were isolated for the first time from adipose tissue suspension extracted from human liposuction. Only a small amount of adipose-derived mesenchymal stem cells can be extracted in a short period of time to cultivate a large number of stem cells for transplantation . [0003] Clinically, the donors and recipients of cell transplantation usually ha...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 陈海佳葛啸虎王小燕张文祺姜交华
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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