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Method for obtaining tissue-engineered cartilage by directional induction of bone marrow mesenchymal stem cells

A bone marrow mesenchymal, directional induction technology, applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., to achieve the effect of inhibiting hypertrophy in vivo, stable cartilage phenotype, and good general shape

Active Publication Date: 2017-06-16
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

However, how to inhibit the hypertrophy and ossification of BMSCs in vivo and maintain the continuous and stable cartilage differentiation is still a bottleneck in clinical application.

Method used

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  • Method for obtaining tissue-engineered cartilage by directional induction of bone marrow mesenchymal stem cells
  • Method for obtaining tissue-engineered cartilage by directional induction of bone marrow mesenchymal stem cells
  • Method for obtaining tissue-engineered cartilage by directional induction of bone marrow mesenchymal stem cells

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Embodiment Construction

[0035] The present invention will be described in detail below in combination with specific embodiments.

[0036] The method for obtaining tissue-engineered cartilage by directional induction of bone marrow mesenchymal stem cells involved in the present invention is realized by the following steps:

[0037] Step 1: Preparation of large aggregates of chondrocytes and extracellular matrix (cell bricks):

[0038] (1) Isolation of chondrocytes, membrane-forming culture:

[0039] The ear cartilage of 1-month-old New Zealand white rabbits was cut into small pieces and treated with 0.2% type II collagenase (Gibco, USA) solution at 37 ○ C for 12 hours, after that, take the single cell suspension and centrifuge at 1000rpm for 5min in a 15ml centrifuge tube;

[0040] Discard the supernatant, wash once with PBS, centrifuge at 1000rpm for 5min, resuspend the cells with film-forming inducing solution, and dilute the cells in a 6.5×10 5 cells / cm 2 The concentration was planted in a 6-we...

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Abstract

The invention relates to a method for obtaining tissue engineering cartilage through directionally inducing bone-marrow mesenchymal stem cells (BMSCs). At present, a large amount of exogenous growth factors and culture in vitro are needed before culturing the BMSCs and the cartilage cells in vivo to promote the differentiation of the BMSCs towards the cartilage cells. The method comprises the following steps of: mixing the sub-cultured P3-generation BMSCs, the cartilage cells and a large extracellular matrix aggregate (cell brick), resuspending the mixture into a platelet rich plasma to prepare a cell compound of the sub-cultured P3-generation BMSCs, the cell brick and the platelet rich plasma, injecting the compound into the skin on the back of a nude mice when the compound condenses gradually, and culturing to obtain the new tissue engineering cartilage. The completely biological and injectable tissue engineering cartilage is obtained on the basis of coculturing the BMSCs and the cartilage cells. The differentiation of the BMSCs towards the cartilage cells can be kept stably, the hypertrophy of the tissue engineering cartilage in vivo can be inhibited effectively, and the defects in the background art can be overcome.

Description

technical field [0001] The invention relates to a method for constructing tissue engineered cartilage, in particular to a method for obtaining tissue engineered cartilage by directional induction of bone marrow mesenchymal stem cells. Background technique [0002] How to construct an injectable tissue-engineered cartilage with a sustained and stable cartilage phenotype is a major challenge for clinical cartilage defect repair. Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into cartilage. When BMSCs are co-cultured with chondrocytes, the chondrogenic differentiation of BMSCs will be promoted. The current method to promote the differentiation of BMSCs into cartilage requires a large number of exogenous growth factors and in vitro culture operations before in vivo culture. However, how to inhibit the hypertrophy and ossification of BMSCs in vivo and maintain the continuous and stable cartilage differentiation is still a bottleneck in clinical ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775A61L27/38
Inventor 吴炜巴睿恺赵铱民
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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