Method for constructing cartilage by inducing human bone marrow stroma stem cell in vitro

A stem cell and cartilage technology, which is applied in the fields of medicine and biomedical engineering, can solve the problems of uneven distribution, affecting the in vitro cartilage construction and clinical application of human BMSCs, and difficulty in cartilage, and achieves the effect of simple preparation.

Active Publication Date: 2011-09-07
SHANGHAI TISSUE ENG LIFE SCI
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Problems solved by technology

[0009] However, human BMSCs are fundamentally different from animal cells, including cell proliferation, differentiation, cell surface antigens, and ability to respond to exogenous inducers. Although many authors have tried to use human BMSCs to construct cartilage tissue, there are The key problems in the following aspects: 1. Poor induction of growth factors; 2. Lack of specificity in the induction of growth factors; 3. Less synthesis and secretion of cartilage-specific matrix; 4. Poor mechanical strength of tissue constructed in vitro ;5. Uneven distribution inside and outside the building tissue; 6. It is difficult to build cartilage with special shapes (such as auricles, nose wings, etc.)
These difficulties seriously affect the in vitro cartilage construction and clinical application of human BMSCs

Method used

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  • Method for constructing cartilage by inducing human bone marrow stroma stem cell in vitro
  • Method for constructing cartilage by inducing human bone marrow stroma stem cell in vitro
  • Method for constructing cartilage by inducing human bone marrow stroma stem cell in vitro

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preparation example Construction

[0059] The preparation method of the tissue engineered cartilage of the present invention is simple and convenient. A certain amount of hBMSCs is mixed with a pharmaceutically acceptable biodegradable material, and then induced in vitro.

[0060] In the process of constructing cartilage in the tissue engineered cartilage graft of the present invention, TGF-β 1 , IGF-I and dexamethasone and other growth factors can be induced in vitro, and various other cytokines or growth factors can also be preferably added or compounded, such as BMP-2, CDMP, etc., so as to accelerate the induction process and improve the synthesis capacity of extracellular matrix, etc. .

[0061] A certain dose of TGF-β1, IGF-I and dexamethasone can effectively induce chondrogenic differentiation of hBMSCs, but the dose of porcine BMSCs can not achieve good induction efficiency when applied to hBMSCs, and the application dose of growth factors must be increased. In a preferred example, TGF-β 1 The concentr...

Embodiment 1

[0079] Acquisition and culture of hBMSCs

[0080] The cells are taken from the human sternum or ilium, and the age of the donor is 8-36 years old, in good health, without malignant tumors, infectious diseases and blood system diseases.

[0081] (1) Material collection: After satisfactory anesthesia, drape was routinely disinfected and punctured at the sternum or ilium with a 16-gauge needle, and 6-8 ml of bone marrow was extracted and placed in a heparinized centrifuge tube.

[0082] (2) Separation of nucleated cells (taking density gradient centrifugation as an example): the extracted bone marrow was aspirated several times successively with 5ml and 1ml empty needles, transferred to another centrifuge tube, and a small amount of serum-free DMEM culture medium was added. Mix well, centrifuge at 3000rpm for 10 minutes, gently suck off the fat and most of the supernatant, pay attention not to stir the sediment below, re-oscillate and mix, and add freshly prepared Percol separati...

Embodiment 2

[0087] Fabrication of PGA three-dimensional scaffold

[0088] Non-woven PGA fibers were purchased from Albany Company (Albany, NY, USA) and stored in vacuum. The fiber diameter is 13-15 μm. Accurately weigh the PGA fiber to 6mg / block, and use a special mold to press it into a small cylinder with a diameter of 5mm and a thickness of 2mm. After the shape is fixed, the material support is completely immersed in 75% ethanol for 30 minutes for disinfection. Rinse with PBS 3 times, then soak in DMEM medium containing 10% fetal bovine serum for 10 minutes, blot dry and prepare to inoculate cells.

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Abstract

The present invention discloses a tissue-engineered cartilage grafting materials, it includes: (a) human bone marrow stem cells (human bone marrow stem cells, hBMSCs); (b) pharmacy acceptable, degradable materials with good biocompatibility. The tissue-engineered cartilages in the invention can effectively grafting treat cartilage defection. The present invention also provides the specific construction method of tissue-engineered cartilage and multiple uses thereof.

Description

technical field [0001] The invention relates to the fields of medicine and biomedical engineering, and more specifically relates to constructing tissue-engineered cartilage by using human bone marrow stromal stem cells (hBMSCs) and its preparation method and application. Background technique [0002] Trauma, infection, tumor or osteochondritis and other diseases can cause cartilage damage. Due to the lack of stem cells and blood supply in cartilage tissue, the self-repair ability after injury is very low, even if there is a small amount of self-repair in some areas (such as small Area articular cartilage can be regenerated after injury), but the repaired tissue is often a composite of fibrous tissue and cartilage tissue, which lacks complete chemical components and sufficient mechanical properties, and cannot perform normal cartilage functions. And some scaffolding cartilage, such as auricular cartilage, completely loses its ability to repair after injury or infection, resul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/44C12N5/077
Inventor 曹谊林周广东刘天一刘伟崔磊
Owner SHANGHAI TISSUE ENG LIFE SCI
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