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Cell mixing method for constructing tissue engineering cartilage

A chondrocyte and tissue engineering technology, applied in the medical field, can solve problems such as poor long-term effect of chondrocytes, and achieve the effects of good effect, high conversion rate and simple method.

Inactive Publication Date: 2007-09-19
尹飞 +1
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AI Technical Summary

Problems solved by technology

MSCs come from a wide range of sources, especially the source of bone marrow is sufficient, it is convenient to obtain materials, there is little damage to the donor site, and there are few postoperative complications. There are still many problems to be solved in terms of directional induction, and many induced chondrocytes degenerate after leaving the induction environment after transplantation, resulting in poor long-term effect of transplantation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1) After the cartilage tissue is obtained from the donor, primary chondrocytes are obtained by type II collagenase digestion.

[0018] 2) The bone marrow of the donor is extracted, and the mesenchymal stem cells (MSCs) are purified by density gradient centrifugation.

[0019] 3) Mixed culture

[0020] After counting the obtained chondrocytes and mesenchymal stem cells (MSCs), the two cells were mixed in proportion so that the proportion of chondrocytes was 70%, and then inoculated into 24-well culture plates, which were pre-coated with Polylysine (PLL) slides allow cells to grow on slides. The culture medium was changed every other day (IMDM culture medium, supplemented with 15% newborn bovine serum, 50 mg / L vitamin C, 100 U / mL penicillin, 100 U / ml streptomycin, pH7.2). After 10 days of mixed culture, the coverslips were removed for detection.

[0021] 4) Cell morphological changes and immunocytochemical findings after mixed cell culture

[0022] When chondrocytes a...

Embodiment 2

[0026] 1) After the cartilage tissue is obtained from the donor, primary chondrocytes are obtained by type II collagenase digestion.

[0027] 2) The bone marrow of the donor is extracted, and the mesenchymal stem cells (MSCs) are purified by density gradient centrifugation.

[0028] 3) Construction of artificial cartilage by mixed cell centrifugation

[0029] After counting the obtained chondrocytes and mesenchymal stem cells (MSCs), the two kinds of cells were mixed in proportion so that the proportion of chondrocytes was 80%, and then 1.5×10 5 Move the mixed cells into a 15ml centrifuge tube at a cell density of / ml, and centrifuge the cells slightly to make them accumulate at the bottom of the centrifuge tube. It can be seen that small cell clusters are formed. Continue to culture in vitro, and change the culture medium every other day (IMDM culture medium, supplemented with 15% Newborn bovine serum, 50mg / L vitamin C, penicillin 100U / mL, streptomycin 100U / ml, pH7.2). Afte...

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Abstract

A method for construction of mixed cells of tissue engineering cartilage, relates to a method improvment of cartilage tissue engineering. The method comprises, 1 primary chondrocyte being obtained using II collagenase digestion method after cartilage tissue is obtained from donor; 2 extracting doner bone marrow and purifying mesenchymal stem cells by density gradient centrifugation method; 3 mixed culture. The effects thereof are abundant source of cell, simple method and high conversion of MSCs into cartilage.

Description

technical field [0001] The invention belongs to the field of medical technology and relates to an improvement of a cartilage tissue engineering method. Background technique [0002] The shortage of seed cells is one of the problems faced by cartilage tissue engineering. At present, the relatively mature autologous chondrocyte transplantation needs about 4.5×10 6 cells, and the artificial cartilage required for animal experiments also needs to be 2.0×10 6 Cell. In order to solve the problem of insufficient seed cells, scholars from all over the world have carried out a lot of research, such as inducing the transformation of bone marrow mesenchymal cells into chondrocytes, inducing the transformation of embryonic stem cells into chondrocytes, and studying the immortalization of chondrocytes. The use of primary chondrocytes has not been reported yet. [0003] Autologous and allogeneic chondrocytes are currently the main cell sources for cartilage tissue engineering, but the ...

Claims

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Application Information

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IPC IPC(8): A61L27/38
Inventor 尹飞郭丽
Owner 尹飞
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