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Construction method for human amniotic epithelial cells and silk fibroin support complexus

An amniotic epithelial cell and silk fibroin technology is applied in the field of constructing complexes of human amniotic epithelial cells in vitro with silk fibroin scaffolds, which can solve the problems of lack of spontaneous regeneration of neuron axons and hinder the self-repair function of CNS, and achieve Proliferative effect

Inactive Publication Date: 2013-04-17
吴卫江
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Problems solved by technology

[0002] The treatment and functional reconstruction of the central nervous system (central nervous system, CNS) after injury is still a major problem in the biomedical field. The traditional concept is that neurons and axons after injury have almost no spontaneous regeneration ability, which hinders the self-regeneration of the CNS. repair function

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  • Construction method for human amniotic epithelial cells and silk fibroin support complexus
  • Construction method for human amniotic epithelial cells and silk fibroin support complexus
  • Construction method for human amniotic epithelial cells and silk fibroin support complexus

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Embodiment Construction

[0015] Main experimental instruments: inverted phase contrast microscope CK-40 (Japan Olympus), fluorescence microscope BX-51 (Japan Olympus), digital imaging system DP-50 (Japan Olympus), CO2 incubator (SPX Corporation), flow cytometer ( Beckman-Coulter Company of the United States), centrifuge TDL-5 (Shanghai Anchan Scientific Instrument Factory), constant temperature cryostat CM1900 (Leica, Germany), petri dish, culture plate (Corning Company of the United States), ultra-clean workbench (Suzhou Su Jing Antai Company), Deep Low Temperature Refrigerator MDF-382E (SANYO Corporation),

[0016] Ultraviolet spectrophotometer (Beijing Opter). Main materials Reagents: RPMI1640 (Gibco Company), fetal bovine serum (U.S. Hyclone Company), trypsin (U.S. Sigma Company), penicillin, streptomycin (Gibco / BRL), EDTA (Sigma), tar violet (Shanghai Kaishi Technology Co., Ltd. ), PE or FITC-labeled mouse anti-human CD29, CD34, CD44, CD45, CD49d, HLA-DR monoclonal antibody (BD Company, USA), N2...

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Abstract

The invention provides a method for constructing amniotic epithelial cells and silk fibroin support complexus in vitro, and the method comprises the following steps of: isolated culture of amniotic epithelial cells; purification of primary cells; identification of primary cells; amniotic cell inoculation on silk fibroin support for space cultivation; and result observation through a scanning electron microscope. According to the method, after human cesarean delivery, wasted amniotic membrane is taken as the material, the amniotic epithelial cells are obtained for cultivation in vitro, and cell purification is carried out under certain conditions, the purified cells and the silk fibroin support are used for constructing a tissue engineering material, thus building a good space growing environment for cells. The invention provides a new way for curing functional repair of all traumatic diseases of the central nervous system by using amniotic membrane cell transplantation in clinical application in the near future.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing complexes of human amniotic epithelial cells and silk fibroin scaffolds in vitro. Background technique [0002] The treatment and functional reconstruction of the central nervous system (central nervous system, CNS) after injury is still a major problem in the biomedical field. The traditional concept is that neurons and axons after injury have almost no spontaneous regeneration ability, which hinders the self-regeneration of the CNS. Repair function. In the 1980s, Aguayo et al. found that CNS axons can be regenerated, but the microenvironment of the central nervous system is not conducive to its regeneration. Therefore, effective external intervention must be carried out to stimulate residual neurons or perform cell transplantation to induce damage. Regenerate nerve axons, restore the transmission of nerve impulses at the damaged site, and finally achieve ...

Claims

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Application Information

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IPC IPC(8): C12N5/073
Inventor 徐杰房文峰朱爱华
Owner 吴卫江
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