Construction for tissue engineering cartilage modified by transforming growth factor beta2 gene

A transforming growth factor and tissue engineering technology, applied in genetic engineering, cells modified by introducing foreign genetic material, tissue culture, etc., can solve the problems of slow metabolism, complicated differentiation and limited sources in in vitro culture, and achieve multiple differentiation potentials , Strong proliferative ability, less trauma to the donor

Inactive Publication Date: 2008-03-26
PEKING UNIV THIRD HOSPITAL
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Problems solved by technology

The isolation and culture of chondrocytes is relatively simple, but the source of chondrocytes is limited, the metabolism in vitro culture is slow, the in vitro expansion is prone to reverse differentiation, and the phenotype (especially type II collagen) is difficult to maintain; the acquisition of BMSCs often requires local anesthesia or dura mater External anesthesia, and the source is limited, the number of stem cells is relatively small
Embryonic stem cells have full differentiation potential, can proliferate indefinitely and differentiate into more than 200 cell types in the whole body, and are expected to form any tissue or organ in the body, but it is extremely complicated to control the differentiation of ESCs cells into specific cells, and the use of ESCs involves biological Safety and Ethical Issues

Method used

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  • Construction for tissue engineering cartilage modified by transforming growth factor beta2 gene

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Embodiment

[0021] Example, In Vivo Construction of Tissue Engineered Cartilage Modified with Transforming Growth Factor beta2 Gene Based on Adipose-derived Adult Stem Cells

[0022] (1) Isolation, culture and expansion of adipose-derived adult stem cells (human adipose derived adult stem cells, hADSCs): A small amount of subcutaneous fat was collected from a small amount of surgical patients after obtaining consent (the patient was a male, aged 56 years, a patient with old femoral neck fracture , underwent total hip replacement), washed three times with PBS to remove visible blood vessels, washed three times with PBS, cut the fat block as much as possible with scissors, put it in a 50ml centrifuge tube, and added 5 times the volume of 0.1% type I collagenase for shaking digestion After 45 minutes, add DMEM containing 10% FBS to stop the digestion, centrifuge at 1200g for 10 minutes, remove the supernatant and lipid droplets, and filter the obtained cell mass with a filter (100 μm) after s...

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Abstract

The present invention discloses process of constructing transforming growth factor beta2 gene modified tissue engineering cartilage. The process includes the separation, culturing and proliferation of adult stem cell from human fat, the transfection of type 5 copying defect adenovirus to the adult stem cell from human fat, the compound of the transfected cell and the polyhydroxy acetic acid / calcium alginate composite as the carrier material, and the formation of in vivo tissue engineering cartilage. Experiment shows that it is possible to form in vivo tissue engineering cartilage through inoculating adult stem cell from human fat to the polyhydroxy acetic acid / calcium alginate composite, and this shows the application foreground of gene reinforced tissue engineering in repairing cartilage defect.

Description

technical field [0001] The present invention relates to the fields of medicine and biomedical engineering, in particular to the field of gene-enhanced tissue engineering. Background technique [0002] Joint pain caused by cartilage loss caused by osteoarthritis or trauma is an important cause of disability in middle-aged and elderly people. The ability of articular cartilage to repair itself is very limited. If it is not treated effectively after injury, it will often cause the degeneration of articular cartilage, and eventually lead to osteoarthritis, with symptoms such as pain, joint stiffness and dysfunction. Commonly used treatment methods include arthroscopic grinding, drilling, microfracture, autologous or allogeneic cartilage transplantation, autologous periosteum or perichondrium transplantation, and prosthetic replacement surgery. However, these traditional treatment options often cannot generate hyaline cartilage and cannot To meet the mechanical requirements of a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N15/12C12N15/861C12N5/10C12N11/04A61L27/00C12N5/0775
Inventor 靳小兵娄思权孙永生
Owner PEKING UNIV THIRD HOSPITAL
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