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Human adipose sub-totipotent stem cell isolated-culture method and stem cell bank establishing method

A technology of subpluripotent stem cells and totipotent stem cells, applied in the field of stem cells, can solve problems such as unfavorable preservation and reduced survival rate of stem cells, and achieve the effects of preventing the introduction of pollution, good cell activity, and uniform tissue digestion.

Inactive Publication Date: 2016-12-07
GENESIS STEMCELL REGENERATIVE MEDICINE ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method is simple, the survival rate of stem cells is greatly reduced, which is not conducive to long-term storage

Method used

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  • Human adipose sub-totipotent stem cell isolated-culture method and stem cell bank establishing method
  • Human adipose sub-totipotent stem cell isolated-culture method and stem cell bank establishing method
  • Human adipose sub-totipotent stem cell isolated-culture method and stem cell bank establishing method

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Experimental program
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Embodiment 1

[0035] A method for separating and culturing human adipose subtotipotent stem cells and a method for constructing a stem cell bank, comprising the following steps: collecting human adipose tissue, isolating and obtaining subtotipotent stem cells, culturing and expanding subtotipotent stem cells, freezing subtotipotent stem cells, product quality control, Stem cell bank building and annual inspection of stem cell recovery; the specific steps are as follows:

[0036] (1) Collection of human adipose tissue: HBV antigen, anti-HCV antibody, anti-HIV antibody, anti-Treponema pallidum antibody, ALT, mycoplasma test items are all negative, no history of blood system disease, immune system disease history, nervous system disease history, Donors with a medical history of the endocrine system; the adipose tissue was obtained from the superficial layer of the abdomen, thigh root, and back through tumescent liposuction in a professional hospital or beauty institution; the separated adipose ...

Embodiment 2

[0046] A method for separating and culturing human adipose subtotipotent stem cells and a method for constructing a stem cell bank, comprising the following steps: collecting human adipose tissue, isolating and obtaining subtotipotent stem cells, culturing and expanding subtotipotent stem cells, freezing subtotipotent stem cells, product quality control, Stem cell bank building and annual inspection of stem cell recovery; the specific steps are as follows:

[0047] (1) Collection of human adipose tissue: HBV antigen, anti-HCV antibody, anti-HIV antibody, anti-Treponema pallidum antibody, ALT, mycoplasma test items are all negative, no history of blood system disease, immune system disease history, nervous system disease history, Donors with a medical history of the endocrine system; the adipose tissue was obtained from the superficial layer of the abdomen, thigh root, and back through tumescent liposuction in a professional hospital or beauty institution; the separated adipose ...

Embodiment 3

[0057] A method for separating and culturing human adipose subtotipotent stem cells and a method for constructing a stem cell bank, comprising the following steps: collecting human adipose tissue, isolating and obtaining subtotipotent stem cells, culturing and expanding subtotipotent stem cells, freezing subtotipotent stem cells, product quality control, Stem cell bank building and annual inspection of stem cell recovery; the specific steps are as follows:

[0058] (1) Collection of human adipose tissue: HBV antigen, anti-HCV antibody, anti-HIV antibody, anti-Treponema pallidum antibody, ALT, mycoplasma test items are all negative, no history of blood system disease, immune system disease history, nervous system disease history, Donors with a medical history of the endocrine system; the adipose tissue was obtained from the superficial layer of the abdomen, thigh root, and back through tumescent liposuction in a professional hospital or beauty institution; the separated adipose ...

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PUM

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Abstract

The invention discloses a human adipose sub-totipotent stem cell isolated-culture method and a stem cell bank establishing method. The human adipose sub-totipotent stem cell isolated-culture method includes the following steps of human adipose tissue collection, sub-totipotent stem cell separation and obtaining, sub-totipotent stem cell culture and proliferation, sub-totipotent stem cell cryopreservation, product quality control, stem cell bank establishing and stem cell revivifying and annual inspection. Mixed collagenase used by the method can effectively separate sub-totipotent stem cells from adipose tissues, is homogenous in tissue digestion, no obvious tissue residual is produced, the cell activity is good, wall adherence is quick, and the homogeneity is good. Cryopreservation liquid adopted by the method can effectively maintain the cell activity, a gradient cooling box is adopted to cryopreserve cells, the cell activity is not affected, and the human adipose sub-totipotent stem cell isolated-culture method is simple in operation, convenient and high in efficiency.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a method for isolating and culturing human adipose subtotipotent stem cells and a method for constructing a stem cell bank. Background technique [0002] Stem cells are a type of cells with self-renewal and multiple differentiation potentials. According to different differentiation potentials, stem cells can be divided into totipotent stem cells, sub-totipotent stem cells, pluripotent stem cells and unipotent stem cells. Sub-totipotent stem cells are a type of differentiation potential second only to Primitive stem cell subsets of totipotent stem cells are at the top of the stem cell hierarchy and are mainly derived from mesenchymal tissues that exist widely in the human body, especially bone marrow, fat, umbilical cord, and placenta, etc. Subtotipotent stem cells are characterized by their epithelial-like morphology , Flk1+CD73+CD90+CD105+CD44+ phenotype, no tumorigenicity, and the cel...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C40B50/06A01N1/02
CPCC12N5/0667A01N1/0221C12N2509/00C40B50/06
Inventor 王军霞殷鉴强刘定生谢再东
Owner GENESIS STEMCELL REGENERATIVE MEDICINE ENG CO LTD
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