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Tissue engineering cartilage construction method using bone matrix gelatin

A technology of tissue engineering and bone matrix, applied in tissue culture, biochemical equipment and methods, medical science, etc., can solve the problems of short maintenance time and a certain distance for permanent functional reconstruction, and achieve strong bone induction and good biological compatibility effect

Inactive Publication Date: 2007-01-31
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently developed tissue engineered cartilage can only replace part of the function of the defective tissue, and the maintenance time is short, and there is still a certain distance from permanent functional reconstruction.

Method used

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  • Tissue engineering cartilage construction method using bone matrix gelatin
  • Tissue engineering cartilage construction method using bone matrix gelatin
  • Tissue engineering cartilage construction method using bone matrix gelatin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1) Seed cell acquisition

[0035] Isolation and culture of chondrocytes

[0036] ①Materials: 1 New Zealand dead rabbit about 4 weeks old was born, depilated with 8% sodium sulfide by mass percentage, washed with running water, soaked in 0.1% bromogeramine for 20-30 minutes; cut off the proximal humerus by aseptic operation, The proximal and distal ends of the femur and the proximal end of the tibia were placed in a container containing D-Hanks' solution containing 100 units / ml of P / S each, and all soft tissues were removed, and the cartilage of each articular surface was sliced ​​off, and placed in a container containing P / S each of 100 units / ml of D-Hanks' solution, then suck out the liquid and discard it, digest it with 10ml hyaluronidase with a mass percentage concentration of 0.05% at room temperature for 3 minutes, discard the enzyme solution, and use After washing with D-Hanks' solution containing 100 units / ml of P / S each, cut the cartilage slices into 1mm 3 sma...

Embodiment 2

[0051] 1) Seed cell acquisition

[0052] Isolation and culture of chondrocytes

[0053] Separation and culture of stromal stem cells to induce chondrocytes:

[0054] ①Using density gradient centrifugation and adhesion separation to obtain bone marrow mesenchymal stem cells: take New Zealand purebred rabbits about four months old, extract bone marrow and slowly inject it on the surface of 70% Percoll cell separation medium at a volume ratio of 1:1, at 1500 Spin centrifuge for 30 minutes, absorb the liquid layer containing bone marrow mesenchymal stem cells between the upper layer and the middle layer, and fill the bone marrow mesenchymal stem cells with DMEM culture solution containing 15% volume ratio of calf serum and P / S 100 units / ml each. Stem cells were made into a single cell suspension, and the nucleated cells were counted and inoculated in culture flasks for monolayer culture. The first 5 days were replaced every 3 days with calf serum containing 15% volume ratio and 1...

Embodiment 3

[0067] 1) Seed cell acquisition

[0068] Isolation and culture of chondrocytes

[0069] Separation and culture of stromal stem cells to induce chondrocytes:

[0070] ①Using density gradient centrifugation and adhesion separation to obtain bone marrow mesenchymal stem cells: Bone marrow puncture was performed on adults, and the bone marrow was extracted and injected slowly on the surface of 70% Percoll cell separation medium at a volume ratio of 1:1, and centrifuged at 1500 rpm for 30 minutes , absorb the liquid layer containing bone marrow mesenchymal stem cells between the upper layer and the middle layer, and prepare the bone marrow mesenchymal stem cells with DMEM culture medium containing 15% volume ratio of calf serum and 100 units / ml of P / S Single cell suspension, count the nucleated cells and inoculate in a culture bottle for monolayer culture, the first 5 days, then every 3 days, replace with calf serum containing 15% volume ratio and P / S 100 units / ml each DMEM cultu...

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Abstract

A tissue engineering cartilage construction method using bone matrix gelatin which comprises, using cartilage cell for isolated culture or base material stem cell for isolated culture to evoke cartilage cell i.e. seed cell, fetching New Zealand rabbit or human embryon os longum and metaphysic trabecular bone to construct bone matrix gel BMG, inoculating the seed cells harvested through isolated culture onto cortex bone or cancellous bone BMG for extracorporal culture.

Description

technical field [0001] The invention belongs to the reconstruction of cartilage tissue in vitro, in particular to a method for constructing tissue engineered cartilage with bone matrix gel. Background technique [0002] At present, the progress of tissue engineering research at home and abroad has opened up a new way for the research and treatment of cartilage diseases, and it is one of the important means for the transformation of future medicine from "replacement" to "reconstruction". Its development prospect is encouraging. With the rapid development of molecular biology and cell biology, the basic theoretical research on various diseases has not only been done at the level of whole animals, isolated organs and tissues, but people have increasingly realized that from the molecular and cellular levels, especially from the living The study of cell morphology and function from the perspective of tissue cells is of great significance to the study of the etiology, pathogenesis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06C12N5/08A61L27/38A61L27/22
Inventor 曹峻岭宋红星李思远孙健谢龙尹战海师钟丽
Owner XI AN JIAOTONG UNIV
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