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Method for preparing human adipose tissue-derived stromal cells

A technology of mesenchymal stem cells and fat, which is applied in the field of preparation of human adipose-derived mesenchymal stem cells, can solve the problems of nausea in patients, consume a lot of manpower and material resources, and residual cryopreservation solution, and achieve less cell damage, better cryopreservation effect, and less side effects Effect

Inactive Publication Date: 2014-06-11
青岛瑞思科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional preparation method has the following disadvantages: 1. The pre-cell culture and cryopreservation recovery process consume a lot of manpower and material resources; 2. After cell recovery, it takes a long time from recovery to full recovery of activity, and the clinical effect in a short period of time is worrying. ;3. After resuscitation, a small amount of cryopreserved fluid remains, which cannot be ruled out in a short time, and will cause adverse reactions such as nausea, vomiting, and fever in patients.

Method used

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  • Method for preparing human adipose tissue-derived stromal cells
  • Method for preparing human adipose tissue-derived stromal cells
  • Method for preparing human adipose tissue-derived stromal cells

Examples

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Embodiment 1

[0029] The preparation method of human adipose-derived mesenchymal stem cells in this embodiment includes the following specific steps:

[0030] (1) Collection of fat: the donors are tested for human-specific viruses including HIV, HBV, HCV, HTLV, EBV, CMV, etc., and tested for Treponema pallidum infection. After passing the test, the fat is collected in a sterile place; Liposuction can be used, and the liposuction sites include: waist, abdomen, buttocks, legs, etc., or surgical methods can be used to collect abdominal or pelvic fat, and the amount of fat should be more than 10ml; after collection, put the fat in the preservation solution , stored at a constant temperature of 2-8°C; the fat preservation solution is composed of DMEM medium + 200 μg / ml penicillin + 200 μg / ml streptomycin + 5 μg / ml amphotericin;

[0031] (2) Transportation of fat: put the fat in the preservation solution, store it in the vaccine box at a constant temperature of 2-8°C, and send it to the laborator...

Embodiment 2

[0040] Identification of adipose-derived mesenchymal stem cells

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Abstract

The invention discloses a method for preparing human adipose tissue-derived stromal cells. The method specifically comprises steps of acquiring, transporting and handing over fat, preparing fat microblocks, carrying out cryopreservation and anabiosis on the fat microblocks, carrying out SVF (Stromal Vascular Fraction) extraction, and carrying out primary culture and subculture. The method is simple and convenient in process, low in cost, good in cryopreservation effect, small in cell damage, small in side effect and harmless to human bodies as the residues of a cryopreservation liquid in cells is reduced to the maximum extent, and the prepared human adipose tissue-derived stromal cells can be applied to clinical treatment of various diseases.

Description

[0001] technical field [0002] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of human adipose-derived mesenchymal stem cells. [0003] Background technique [0004] Adipose tissue contains a type of cell with multi-lineage differentiation potential, that is, adipose-derived mesenchymal stem cell (AD-MSC), its biological properties are similar to bone marrow mesenchymal stem cells, and it can differentiate into fat, bone, etc. , cartilage, muscle, endothelial, hematopoietic, liver, islet and nerve cells. [0005] Adipose tissue is rich in reserves in the human body, easy to obtain and less traumatic, and has broad application prospects in tissue engineering, organ repair, gene therapy, etc. Therefore, adipose stem cells have become a hot spot in the field of stem cells and have broad clinical applications prospect. In January 2012, Cuepistem, an autologous adipose stem cell drug approved by the Korean FDA, was succe...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 许评张炳强宋岗陈立材史光军于丽
Owner 青岛瑞思科生物科技有限公司
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