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Procuring and separating culture method of human adipose-derived stem cells and construction method of pool of stem cells

A human adipose stem cell, separation and culture technology, which is applied in the field of obtaining, separating and culturing human adipose stem cells and the construction of a stem cell bank, can solve the problems of insufficient configuration of the adipose stem cell bank, slow proliferation of adipose stem cells, serious cell differentiation, etc., and achieve adipose tissue. Complete effect, inhibition of stem cell differentiation, and high stem cell purity

Inactive Publication Date: 2018-05-01
北京臻溪谷医学研究中心(有限合伙)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the existing isolation and culture of human adipose stem cells have low tissue digestion, slow proliferation of adipose stem cells, severe cell differentiation, and low survival rate during cell culture. In addition, in order to facilitate people to effectively store and use stem cell resources, currently in Worldwide, many stem cell banks have been established. A perfect stem cell bank should have the ability to provide stored healthy stem cells for clinical use anytime and anywhere. Currently, the adipose stem cell bank constructed in the existing technology is used to store oneself, family members and others in For various diseases that need to be treated with adipose-derived stem cell transplantation and related technologies, providing clinically-used adipose-derived stem cells has broad application prospects. future medical needs

Method used

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  • Procuring and separating culture method of human adipose-derived stem cells and construction method of pool of stem cells
  • Procuring and separating culture method of human adipose-derived stem cells and construction method of pool of stem cells
  • Procuring and separating culture method of human adipose-derived stem cells and construction method of pool of stem cells

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Embodiment 1

[0031] Embodiment 1 of the present invention provides a method for obtaining, separating and culturing human adipose stem cells, the method comprising the following steps:

[0032] S1. Collection of human adipose tissue: Collect the adipose tissue of the donor in a sterile environment, put it in 0.9% sodium chloride injection with a volume ratio of 3:1, wash it, remove the necrotic tissue and connective tissue, and remove the adipose tissue. Tissue cut to 0.5mm 3 size;

[0033] S2. Treatment of human adipose tissue: the adipose tissue was collected in a centrifuge tube, centrifuged at 1800 rpm / min for 15 min, the upper part of the fat layer was collected, and washed with PBS;

[0034] S3. Digestion of human adipose stem cells: add an equal volume of digestive enzymes for 20 min at a temperature of 30 °C, add an equal volume of DMEM complete medium containing 10% serum to terminate the digestion, and after standing for 20 min, remove The upper layer was centrifuged at 1200 rp...

Embodiment 2

[0037] Embodiment 2 of the present invention provides a method for obtaining, separating and culturing human adipose stem cells, the method comprising the following steps:

[0038] S1. Collection of human adipose tissue: Collect the adipose tissue of the donor in a sterile environment, put it in 0.9% sodium chloride injection with a volume ratio of 4:1 to wash it, remove the necrotic tissue and connective tissue, and remove the adipose tissue. Tissue cut to 2mm 3 size;

[0039] S2. Treatment of human adipose tissue: the adipose tissue was collected in a centrifuge tube, centrifuged at 2000 rpm / min for 15 min, the upper part of the fat layer was collected, and washed with PBS;

[0040] S3. Digestion of human adipose stem cells: add an equal volume of digestive enzymes for 30 minutes at a temperature of 35°C, add an equal volume of DMEM complete medium containing 10% serum to terminate the digestion, and after standing for 20 minutes, remove the The upper layer was centrifuged...

Embodiment 3

[0043] Embodiment 3 of the present invention provides a method for obtaining, separating and culturing human adipose stem cells, the method comprising the following steps:

[0044] S1. Collection of human adipose tissue: Collect the adipose tissue of the donor in a sterile environment, wash it in 0.9% sodium chloride injection with a volume ratio of 2:1, remove the necrotic tissue and connective tissue, and remove the adipose tissue. Tissue cut to 3.5mm 3 size;

[0045] S2. Treatment of human adipose tissue: the adipose tissue was collected in a centrifuge tube, centrifuged for 15 minutes at 2200 rpm / min, the upper part of the fat layer was collected, and washed with PBS;

[0046] S3. Digestion of human adipose-derived stem cells: add an equal volume of digestive enzymes for 45 min at a temperature of 37°C, add an equal volume of DMEM complete medium containing 10% serum to terminate the digestion, and after standing for 20 min, remove the The upper layer was centrifuged at ...

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Abstract

The invention provides a procuring and separating culture method of human adipose-derived stem cells and a construction method of a pool of stem cells. The procuring and separating culture method comprises the following steps: collecting human fat tissues; treating the human fat tissues; digesting human fat stem cells; and culturing the human fat stem cells. The construction method of the pool ofstem cells comprises the following steps: procuring, separating and culturing the human fat stem cells; numbering the procured human fat stem cells with IDs, recording and procuring donor informationof the human fat stem cells, and linking the ID numbers to the donor information to form a cell information file and storing the cell information file in a database to form the human fat pool of stemcells. The procuring and separating culture method of human adipose-derived stem cells, provided by the invention, not only is more thorough in fat tissue effect, but also is short in culture time. Inthe culture process, a special culture medium can realize quicker cell proliferation, the purity of stem cells is higher, and in the cell culture process, stem cell differentiation is avoided, and inthe cell cryopreservation process, the survival rate is higher.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and particularly relates to a method for obtaining, separating and culturing human adipose stem cells and a method for constructing a stem cell bank. Background technique [0002] Stem cells are a kind of pluripotent cells with self-replication ability, which can differentiate into various functional cells under certain conditions. Stem cells are cells of origin. In short, they are primitive undifferentiated cells with multi-directional differentiation potential and self-replication ability, and are primitive cells that form various tissues and organs of mammals. Adipose-derived stem cells are a type of stem cells with multidirectional budding potential obtained from adipose tissue, which can differentiate into osteoblasts, chondroblasts and adipocytes under specific induction conditions. Zuk first obtained such stem cells with cell therapy potential by isolating human adipose tissu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A01N1/02C40B50/06
CPCA01N1/0221A01N1/0226C12N5/0667C12N2500/12C12N2501/999C12N2509/00C40B50/06
Inventor 曹毓琳刘俊江时兆田白志惠
Owner 北京臻溪谷医学研究中心(有限合伙)
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