Culture method for inducing chondrogenic differentiation of adipose-derived stem cells and culture solution

A technology for adipose stem cells and culture methods, applied in the field of culture methods and culture fluids for inducing chondrogenic differentiation of adipose stem cells, capable of solving problems such as limitations in culturing bone marrow mesenchymal stem cells

Inactive Publication Date: 2018-08-17
HUNAN YUANPIN CELL TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] At present, in the field of research on mesenchymal stem cells, the main target is bone marrow mesenchymal stem cells, and a method of is...

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  • Culture method for inducing chondrogenic differentiation of adipose-derived stem cells and culture solution
  • Culture method for inducing chondrogenic differentiation of adipose-derived stem cells and culture solution
  • Culture method for inducing chondrogenic differentiation of adipose-derived stem cells and culture solution

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Embodiment Construction

[0016] The technical solutions of the present invention are described below in combination with specific embodiments. The experimental materials that are not particularly emphasized in the following examples are conventional experimental materials without special requirements, and are all conventional materials that are easily obtained by those skilled in the art.

[0017] 1. Experimental materials

[0018] Chizhi aldehyde A and Chizhi aldehyde B standard products are self-made, and the purity is above 98%. The chemical information is as follows.

[0019]

[0020] DMEM medium and fetal bovine serum were purchased from Gibco.

[0021] 2. Experimental method

[0022] 1. Isolation and culture of adipose-derived stem cells

[0023] The subcutaneous adipose tissue of healthy volunteers was taken, washed three times with PBS buffer, and the adipose tissue membrane and blood vessels were removed with ophthalmic scissors and ophthalmic forceps and cut into 1mm 3 For fragments o...

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Abstract

The invention discloses a culture method for inducing chondrogenic differentiation of adipose-derived stem cells and a culture solution. The culture method comprises the following steps: (1) taking asubcutaneous fatty tissue, washing the subcutaneous fatty tissue, removing an adipose tissue membrane and blood vessels and shearing the subcutaneous fatty tissue into broken blocks, carrying out enzymic digestion, adding a complete medium, centrifuging to remove adipose cells and lipid droplets, resuspending on the complete medium, inoculating into a culture bottle, culturing for 24 h and then changing the culture solution to remove cells which are not attached to walls, changing the solution every three days, digesting for 3-5 min with 0.25% of pancreatin and 1 mmol/L EDTA at the temperatureof 37 DEG C after cell fusion, stopping digestion of the complete medium, and carrying out passage at the ratio of 1: 2; and (2) inoculating passage cells into the culture bottle which contains the complete medium, culturing for 48h and then changing the medium into an incomplete medium, adding a Sox9 gene activator to continue culturing, and changing the medium every three days. By the providedmethod, chondrogenic differentiation of the adipose-derived stem cells can be induced effectively, the adipose-derived stem cells can be used as seed cells to prepare a large number of chondrocytes tomeet clinic requirements to the chondrocytes.

Description

technical field [0001] The invention belongs to the field of biology, and relates to a culture method and a culture solution for inducing chondrogenic differentiation of adipose stem cells. Background technique [0002] At present, in the field of research on mesenchymal stem cells, the main target is bone marrow mesenchymal stem cells, and the method of isolation and culture has been initially formed, but the mass culture of bone marrow mesenchymal stem cells is limited. Especially in the study of cartilage tissue engineering, due to the difficulty of isolating and culturing bone marrow mesenchymal stem cells, it is limited to become a good source of tissue engineering seed cells. Therefore, recently people have turned their research to adipose-derived stem cells, and found that the cells have strong proliferation and differentiation capabilities, and are easier to isolate and culture. In clinical research, adipose-derived stem cells also have broad application prospects i...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775
CPCC12N5/0655C12N5/0667C12N2501/999C12N2506/1384C12N2509/00
Inventor 田红青
Owner HUNAN YUANPIN CELL TECH CO LTD
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