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Detection method of adenovirus-mediated gene transfection autologous bone marrow mesenchymal stem cells

A bone marrow mesenchymal and gene transfection technology, which is applied in the detection field of adenovirus-mediated gene transfection of autologous bone marrow mesenchymal stem cells, and can solve the problems of influence, biological and mechanical properties defects, etc.

Inactive Publication Date: 2017-01-04
GUILIN MEDICAL UNIVERSITY
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Problems solved by technology

[0009] The purpose of the present invention is to provide a detection method for adenovirus-mediated gene transfection of autologous bone marrow mesenchymal stem cells. The process of proliferation, differentiation and degeneration affects ANFH, and the current cell growth scaffolds have defects in biological and mechanical properties in the process of tissue construction

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  • Detection method of adenovirus-mediated gene transfection autologous bone marrow mesenchymal stem cells
  • Detection method of adenovirus-mediated gene transfection autologous bone marrow mesenchymal stem cells

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[0064] 3. Preparation, biocompatibility testing and mechanical property analysis of allogeneic DBM materials. After the healthy New Zealand white rabbits were sacrificed, the metaphyseal bones of the limbs and cancellous vertebral bodies were collected, and stored at -80°C for 72 hours according to the method of Urist et al. Provided) for 72 hours of decalcification, degreasing in ether and ethanol for 24 hours each, repeated washing and soaking with sterile distilled water until the soaking solution is neutral. At this time, the cancellous bone is white and spongy, which can be compressed and deformed and automatically return to its original shape. After deacidification treatment, the bone blocks were soaked in distilled water for 24 hours, taken out, and dried naturally to make a 4mm×4mm×3mm decalcified bone matrix, sterilized with ethylene oxide, and stored at 4°C for later use. Under an inverted microscope, it was observed that the scaffold material had a good pore struct...

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Abstract

The invention discloses a detection method of adenovirus-mediated gene transfection autologous bone marrow mesenchymal stem cells. The detection method includes: using hBMP-2 genes to transfect bone marrow mesenchymal stem cells BMSCs by the aid of a recombinant defective adenovirus expression carrier Ad-hBMP-2, compositing the transfected cells with a allogeneic decalcified bone DBM stent material in an in-vitro manner, implanting into an ANFH animal model, and performing X-ray detection, transmission electron microscope detection, histological detection, immunohistochemical analysis (semiquantitative) and mechanical performance detection on the repaired femoral head. By the detection method, the conditions and influence factors of the proliferation, directional differentiation and degeneration of the stem cells are illustrated, cytology theoretical support is provided for methods using the stem cells to treat ANFH, and basis and support are provided for the clinical application of the methods using the stem cells to treat the ischemic necrosis of the femoral head by successfully demonstrating the relation between the number, differentiation, degeneration and the like of the bone marrow mesenchymal stem cells and the ischemic necrosis of the femoral head.

Description

technical field [0001] The invention belongs to the field of medical technology, and in particular relates to a detection method for adenovirus-mediated gene transfection of autologous bone marrow mesenchymal stem cells. Background technique [0002] The latest study found that the activity and quantity of bone marrow mesenchymal stem cells in patients with avascular necrosis of the femoral head (ANFH) decreased, suggesting that the cause of the disease may not be the osteoblast itself but the proliferation or proliferation of mesenchymal stem cells. Differentiation is inhibited. Stem cell transplantation also has the phenomenon of cell phenotype variation and loss of osteogenic ability. [0003] Avascular necrosis of femoral head (ANFH) is a common clinical orthopedic disease with a high disability rate. The current treatment measures include core decompression, drilling and bone grafting for early cases, and femoral head replacement for advanced cases. The effects of th...

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Application Information

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IPC IPC(8): C12N15/861C12N15/12C12N5/10C12Q1/06G01N33/68C12Q1/68
CPCC07K14/51C12N5/0663C12Q1/686G01N33/502G01N33/5026G01N33/68C12Q2561/113C12Q2563/107C12Q2545/114C12Q2531/113
Inventor 李强赵美玉李诗鹏茹嘉武成聪陈佳滨宁寅宽蔡伟良石正松
Owner GUILIN MEDICAL UNIVERSITY
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