Method for converting in-vitro activated adipose-derived stem cells into chondroblasts

A technology of adipose stem cells and chondrocytes, which is applied in the field of activating adipose stem cells in vitro and transforming them into primary chondrocytes, can solve the problems of low clinical practicability, hypertrophy and hyperplasia of chondrocytes, and lack of specific surface antigens.

Inactive Publication Date: 2020-08-11
GUANGDONG XIANKANGDA BIOTECH CO LTD
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Problems solved by technology

[0003] The main problems are: (1) There are many types of ASCs surface molecules known at present, but most of them cross with other mesenchymal stem cells, and there is no clear specific surface antigen, so it is difficult to select one of the highly specific surface marker antigens Combined screening; (2) Although many laboratories try to use different materials and techniques to induce ASCs to differentiate into cartilage tissue, they cannot avoid the result of inflammatory reaction between cartilage tissues; (3) In many experimental cultured chondrocytes, the late The experimental results all showed the hypertrophy and hyperplasia of chondrocytes; (4) At present, the induction of ASCs to differentiate into chondrocytes basically adopts the construction of 3D models, but the clinical practicability of this method is not high

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Embodiment Construction

[0024] It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0025] A method for activating adipose stem cells into prochondrocytes in vitro, adipose tissue to be treated according to the present invention can be obtained from a patient or an immunologically acceptable donor, isolating adipose tissue from an adipose sample, and culturing the activated adipose tissue . An "immunologically acceptable donor" is a human having tissue, including adipose tissue. Firstly, the adipose tissue was washed with buffer solution to remove residual blood and tissue fragments. In this example, D-Hanks buffer solution was used to wash the adipose tissue. After leaving it layered, absorb the upper fat cell liquid; and place the fat cell liquid in a box at 0-5 degrees Celsius for 1-2 hours. Hours can increase the activity of adipocyte liquid, then seal and centrifuge, remove the supernatant, an...

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Abstract

The invention discloses a method for converting in-vitro activated adipose-derived stem cells into chondroblasts. The method comprises the following steps of preparing a DMEM culture medium, wherein the DMEM culture medium is prepared from 1% of fetal calf serum, 10 ng / ml of TGF-beta, 100 ng / ml of BMP-6, 6.25 mu g / ml of insulin, 0.1 mu mol / L of dexamethasone, 6.25 mu g / ml of transferrin and 50 mumol / L of ascorbyl phosphate; culturing the adipose-derived stem cells formed into micelles in the DMEM culture medium; and injecting the adipose-derived stem cells cultured for 7-9 days into an articular cavity and converting the adipose-derived stem cells into chondroblasts. The method has the advantages of time saving, simple process control and low rejection of the generated chondroblasts.

Description

technical field [0001] The invention relates to the field of medicine, in particular to a method for activating adipose stem cells in vitro to transform into prochondrocytes. Background technique [0002] Among the mesenchymal stem cells (MSCs) that have been used more so far, bone marrow derived stem cells (BMSCs) have been a research hotspot and have been widely used in scientific research. In 2001, Zuk et al. isolated for the first time multidirectionally differentiated stem cells from adipose tissue suspension extracted by liposuction. Adipose-derived mesenchymal cells (ASCs) are superior to BMSCs because stem cells extracted from adipose tissue have the advantages of easy sampling, large amount of sampling, repeated sampling and rapid cell proliferation. At the same time, ASCs can differentiate into chondrocytes, osteoblasts, adipocytes, nerve cells, etc. in a specific microenvironment. Therefore, ASCs are expected to be good seed cells for tissue engineering. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/0775
CPCC12N5/0655C12N5/0667C12N2500/14C12N2500/25C12N2500/38C12N2501/06C12N2501/15C12N2501/155C12N2501/2302C12N2501/39
Inventor 谢海涛薛卫巍王斌
Owner GUANGDONG XIANKANGDA BIOTECH CO LTD
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