Clinical-level purification, separation, culture amplification and cryopreservation method of fat-derived mesenchymal stem cells

A stem cell and adipose-derived technology, applied in the field of cell biological products, to achieve the effect of increasing cell volume and reducing unit cost

Pending Publication Date: 2020-10-13
深圳市人和生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Based on this, the purpose of the present invention is to solve the problem of clinical-grade production and preparation of adipose-derived mesenchymal stem cells through the improvement of technology and technical methods, meet the needs of adipose-derived mesenchymal stem cells in clinical research, and provide an adipose-derived mesenchymal stem cell. Methods for clinical-grade purification, isolation, culture expansion and cryopreservation of mesenchymal stem cells

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  • Clinical-level purification, separation, culture amplification and cryopreservation method of fat-derived mesenchymal stem cells
  • Clinical-level purification, separation, culture amplification and cryopreservation method of fat-derived mesenchymal stem cells
  • Clinical-level purification, separation, culture amplification and cryopreservation method of fat-derived mesenchymal stem cells

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[0043] The implementations described in the following exemplary examples do not represent all implementations consistent with the present disclosure. Rather, they are merely examples of approaches consistent with aspects of the disclosure as recited in the appended claims.

[0044] The terminology used in the present disclosure is for the purpose of describing particular embodiments only, and is not intended to limit the present disclosure. As used in this disclosure and the appended claims, the singular forms "a", "the", and "the" are intended to include the plural forms as well, unless the context clearly dictates otherwise. It should also be understood that the term "and / or" as used herein refers to and includes any and all possible combinations of one or more of the associated listed items.

[0045] Before implementing the clinical-grade purification and separation, culture expansion and cryopreservation methods of adipose-derived mesenchymal stem cells of the present inv...

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Abstract

The present invention discloses a clinical-level purification, separation, culture amplification and cryopreservation method of fat-derived mesenchymal stem cells. The method comprises the following steps: S1. collecting adipose tissue; S2. Adding a washing liquid to wash the adipose tissue; S3. adding a type I collagenase digestion solution to a culture flask to obtain a fat SVF cell filtrate; S4. centrifuging the adipose SVF cell filtrate to obtain a precipitate, namely, fat SVF; S5. discarding a supernatant to obtain a purified fat SVF precipitate; S6. continuing subculture of the culture flask; S7. performing primary culture to obtain primary cells; S8. digesting and harvesting the primary cells, adding the culture medium and performing resuspension, sampling and counting; S9. centrifuging the cell suspension obtained by S8 again, discarding the supernatant, and culturing the culture flask; S10. when the cell fusion in S9 reaches 80% or more, repeating the S8 and S9, and carrying out digestion and washing to obtain a cell precipitate; S11. Sampling the cell precipitate in S10 for detection; and S12. adding a complete culture solution to the cell precipitate obtained after resuspension in the S11, and after the test result is qualified, transferring the cell precipitate to a cell bank for long-term storage.

Description

technical field [0001] The invention relates to the technical field of cell biological products, in particular to a method for clinical grade purification, isolation, culture expansion and cryopreservation of adipose-derived mesenchymal stem cells. Background technique [0002] Adipose-derived mesenchymal stem cells are a kind of stem cells with multi-differentiation potential isolated and purified from adipose tissue, which can differentiate into cartilage, osteoblast, fat, nerve tissue and other tissues or organs under the action of certain inducing factors. The advantages of adipose-derived mesenchymal stem cells as applied cells are: 1. The source of raw materials is simple and easy to obtain; 2. It has the characteristics of mesenchymal stem cells; 3. It is widely used in the fields of knee joints and medical beauty. Cells with safe quality provide corresponding technical support for clinical research. [0003] Existing technical methods and processes use animal-derive...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A01N1/02
CPCC12N5/0667A01N1/0221C12N2509/00C12N2509/10C12N2500/90
Inventor 王广平王知源
Owner 深圳市人和生物科技有限公司
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