Composition for treating defects of articular cartilage
A technology of articular cartilage and composition, which is applied in the direction of drug combination, bone disease, medical preparations containing active ingredients, etc., and can solve problems such as joint swelling and soreness
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0144] Example 1 Cell isolation, purification and cultivation
[0145] a) Wash adipose tissue (remove blood cells): add an equal amount of normal saline to the fat-containing centrifuge tube, tighten the cap, shake for 3 minutes to fully wash the adipose tissue, then stand still for 3-5 minutes to separate the different phases, and absorb the lower layer of water phase; repeat the above operation three times until the lower layer is relatively clear.
[0146] b) Collagenase digestion: After aspirating and discarding the normal saline, add preheated DMEM containing 0.1% collagenase I equal to the volume of fat, put it in a constant temperature oscillator, 37°C, 200rpm, digest for 1h, shake and centrifuge every 15min Tube for 5-10 seconds (make the fat fully contact with collagenase I).
[0147] c) Collect the precipitate: after digestion, centrifuge at 2000rpm for 10min, discard the digested fat in the upper layer, collect the bottom layer of the two tubes into a new centrifug...
Embodiment 2
[0151] Example 2 Identification of adipose progenitor cells
[0152] The adipose progenitor cells cultured in Example 1 were centrifuged and resuspended; after cell counting, the cell concentration was adjusted to 1×10 8 / L, react with human anti-CD34, CD45, CD29, CD73, CD90, CD105, Actin, CD14, HLA-DR monoclonal antibody at room temperature for 30min respectively, resuspend the cells with PBS, and detect with flow cytometry ( figure 1 ).
[0153] surface antigen
[0154] Conclusion: Analysis of the expression of cell surface antigen markers on adipose progenitor cells by flow cytometry shows that the cells are of high purity.
Embodiment 3
[0155] Example 3 Test of chondrogenic, osteogenic and adipogenic differentiation ability of adipose mesenchymal progenitor cells
[0156] The cells cultured in Example 1 were used as the group of the present invention, and the chondrogenic, osteogenic and adipogenic differentiation abilities were tested. Alcian blue staining shows that the cells cultured in Example 1 have the ability to differentiate to cartilage in vitro after 3-4 weeks of differentiation and culture in the direction of cartilage ( figure 2 A) Alizarin red staining shows that the cells cultured in Example 1 have the ability to differentiate to bone in vitro after 3-4 weeks of cultured in vitro to bone direction differentiation ( figure 2 B); oil red O staining shows that the cells cultured in Example 1 have the ability to differentiate to fat in vitro after 3-4 weeks of culture in the direction of fat differentiation ( figure 2 C).
PUM
| Property | Measurement | Unit |
|---|---|---|
| volume | aaaaa | aaaaa |
| volume | aaaaa | aaaaa |
| molecular weight | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More