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Human fat mesenchymal stem cell bank and construction method thereof

A technique of stromal stem cells and a construction method, which is applied in the field of human adipose-derived mesenchymal stem cell bank and its construction, can solve the problems of inability to extract bone marrow, immune response, and the decline of proliferation and differentiation ability, and achieves easy autologous transplantation, high separation efficiency, high source wide range of effects

Active Publication Date: 2013-05-01
冯文峰
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Problems solved by technology

However, bone marrow-derived mesenchymal stem cells have the following problems: with aging, the number of stem cells decreases significantly, and the ability to proliferate and differentiate significantly declines; the preparation process is not easy to control the quality; transplantation to the allogeneic body may cause immune reactions; and when the material is obtained, it is harmful to the patient. Injury, when the patient has bone marrow disease, it cannot be collected, even if it is a healthy donor, too much bone marrow cannot be extracted
However, the sources of placenta and umbilical cord tissue are limited, and people who have already been born can no longer obtain placenta and umbilical cord, which induces us to look for the ideal source of mesenchymal stem cells

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  • Human fat mesenchymal stem cell bank and construction method thereof

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Embodiment approach

[0032] As a preferred embodiment of the method for constructing a human adipose-derived mesenchymal stem cell bank of the present invention, the serum is autologous serum.

[0033] As a preferred embodiment of the construction method of the human adipose-derived mesenchymal stem cell bank of the present invention, the autologous serum is prepared by the following method: aseptically collect 20 mL of peripheral venous blood, leave it in a 4°C refrigerator for 4h, and place it in a 37°C incubator Incubate for 4 hours, centrifuge at 1500r / min for 5 minutes, absorb the upper serum, filter and sterilize at 0.22μm, aliquot in the ultra-clean workbench, test negative for bacterial culture, and store at -20°C for later use.

[0034] As a preferred embodiment of the method for constructing a human adipose-derived mesenchymal stem cell bank of the present invention, the human adipose-derived mesenchymal stem cell bank is an autologous bank.

[0035] A human adipose-derived mesenchymal s...

Embodiment 1

[0038] The human adipose-derived mesenchymal stem cell bank was constructed by the following method:

[0039] (1) Sterile collection of human body fat: First, ABO / RH blood typing, HLA typing, microbiological testing, HIV, HBV, HCV, TP testing were performed on the adipose-derived mesenchymal stem cell preservers. Negative; then aseptically collect fat, 200 mL of fat can be taken at one time, from which about 1×10 6 mesenchymal stem cells.

[0040] (2) Removal of grease: Cut the obtained adipose tissue with sterile surgical scissors or a knife, the length of which should not exceed 3mm, then wash it repeatedly with an equal volume of PBS solution, let it stand for equal layers, and remove the lower layer of grease / Lipid, upper fraction collected.

[0041] (3) Collagenase digestion: Add 0.075% type Ⅰ collagenase, shake and digest at 37 °C for 30 min, then stop the digestion with DMEM-LG medium containing 10% autologous serum; centrifuge at 280g for 5 min, discard the supernatan...

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Abstract

The invention discloses a human fat mesenchymal stem cell bank and a construction method thereof. The human fat mesenhymal stem cell bank is constructed by the following steps: (1), aseptically acquiring human fat; (2) removing oil and fat; (3), digesting collagenase; (4) performing schizolysis on red blood cells; (5) expanding culture; and (6), cryopreservation. A human fat mesenchymal stem cell is a type of pluripotent stem cell of fat tissue, can induce, differentiate and form cells of tissue types such as fat, bone, cartilage, muscle and the like. Fat tissue is wide in sources and easy for material obtaining, does not relate to ethical issues, is convenient for autograft, and brings less pain to patients; and in addition, compared with the mesenchymal stem cells of sources such as marrow and the like, the fat mesenchymal stem cell is fast in multiplication and high in separation efficiency.

Description

technical field [0001] The present invention relates to a stem cell bank and its construction method, in particular to a human adipose mesenchymal stem cell bank and its construction method. Background technique [0002] Mesenchymal stem cells (MSCs) are important members of the stem cell family, derived from the mesoderm and ectoderm in the early stages of development. Mesenchymal stem cells were originally discovered in the bone marrow, and have attracted increasing attention because of their multi-lineage differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and self-replication. Under specific induction conditions in vivo or in vitro, mesenchymal stem cells can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelial and other tissue cells. It has multi-directional differentiation potential and can be used as an ideal seed cell for the repair of tissue and organ dam...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 冯文峰列浦昌佘志勇方海庆
Owner 冯文峰
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