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Method for preparing matrix-induced autologous bone mesenchymal stem cell composite scaffold

A technology of bone marrow mesenchymal and composite scaffolds, which is applied in the field of preparation of autologous bone marrow mesenchymal stem cell composite scaffolds, can solve the problems of poor detection accuracy and insufficient sensitivity, reduce the number of operations, reduce treatment costs, and facilitate passage and expansion. Increased effect

Inactive Publication Date: 2017-07-25
安徽安龙基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to solve the defects of poor detection accuracy and insufficient sensitivity in the prior art, and provide a method for preparing a matrix-induced autologous bone marrow mesenchymal stem cell composite scaffold to solve the above problems

Method used

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Examples

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Effect test

preparation example Construction

[0037] The invention discloses a preparation method of a matrix-induced autologous bone marrow mesenchymal stem cell composite scaffold. The specific steps are:

[0038] (1) Under sterile conditions, use a 20ml syringe to draw 1ml of heparin sodium saline into the syringe, and then continue to use the syringe to draw 9ml of bone marrow. After removing the needle, add 10ml of heparin sodium saline and bone marrow. In the culture bottle of DMEM high glucose medium, after mixing evenly, obtain the bone marrow suspension;

[0039] (2) Under sterile conditions, take 100-200ml of peripheral blood from the patient, put it in a blood bag, and seal it;

[0040] (3) Divide the patient's peripheral blood into 50ml centrifuge tubes, 45ml per tube, centrifuge at 3000rpm for 30min, take the supernatant, centrifuge at 3000rpm for 30min, remove the precipitate, and 0.22μm pore size filter sterilization to obtain autologous serum, which is packed for use;

[0041] (4) Slowly add the bone mar...

Embodiment 1

[0058] The invention discloses a preparation method of a matrix-induced autologous bone marrow mesenchymal stem cell composite scaffold. The specific steps are:

[0059] (1) Under sterile conditions, use a 20ml syringe to draw 1ml of heparin sodium saline into the syringe, and then continue to use the syringe to draw 9ml of bone marrow. After removing the needle, add 10ml of heparin sodium saline and bone marrow. In the culture bottle of DMEM high glucose medium, after mixing evenly, obtain the bone marrow suspension;

[0060] (2) Under sterile conditions, take 150ml of peripheral blood from the patient, put it in a blood bag, and seal it;

[0061] (3) Divide the patient's peripheral blood into 50ml centrifuge tubes, 45ml per tube, centrifuge at 3000rpm for 30min, take the supernatant, centrifuge at 3000rpm for 30min, remove the precipitate, and 0.22μm pore size filter sterilization to obtain autologous serum, which is packed for use;

[0062] (4) Slowly add the bone marrow ...

Embodiment 2

[0079] The invention discloses a preparation method of a matrix-induced autologous bone marrow mesenchymal stem cell composite scaffold. The specific steps are:

[0080] (1) Under sterile conditions, use a 20ml syringe to draw 1ml of heparin sodium saline into the syringe, and then continue to use the syringe to draw 9ml of bone marrow. After removing the needle, add 10ml of heparin sodium saline and bone marrow. In the culture bottle of DMEM high glucose medium, after mixing evenly, obtain the bone marrow suspension;

[0081] (2) Under sterile conditions, take 150ml of peripheral blood from the patient, put it in a blood bag, and seal it;

[0082] (3) Divide the patient's peripheral blood into 50ml centrifuge tubes, 45ml per tube, centrifuge at 3000rpm for 30min, take the supernatant, centrifuge at 3000rpm for 30min, remove the precipitate, and 0.22μm pore size filter sterilization to obtain autologous serum, which is packed for use;

[0083] (4) Slowly add the bone marrow ...

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Abstract

The invention discloses a method for preparing a matrix-induced autologous bone mesenchymal stem cell composite scaffold. The method specifically comprises the steps of preparing bone marrow suspension; taking peripheral blood of a patient and putting the peripheral blood of the patient in a blood bag; preparing autoserum; obtaining target cells; adding 15 ml DMEM high glucose culture medium to the target cells, inoculating a T25 cell culture bottle with the target cells, and placing the T25 cell culture bottle in an incubator for culture; rinsing the cells twice with PBS solution; adding the DMEM high glucose culture medium into the culture bottle dropwise to stop digestion; obtaining cleaned sediment; inoculating the cell culture bottle with the cells for continuous culture; conducting passage three times; adding trypsin-EDTA solution into the culture bottle; washing the cells three times with PBS; conducting cell resuspension; placing a sterile pig source I / III type collagen membrane in a sterile plastic culture dish, and soaking the membrane in the DMEM high glucose culture medium for one night; and dropping cell suspension onto the pig source I / III type collagen membrane, and conducting culture for 3 days. The method has the advantages of being high in treatment precision and the like.

Description

technical field [0001] The invention relates to the field of tissue engineering articular cartilage injury repair treatment, in particular to a preparation method of matrix-induced autologous bone marrow mesenchymal stem cell composite scaffold. Background technique [0002] Articular cartilage injury is a common clinical disease. The clinical manifestations are joint pain, which is aggravated by weight bearing and exercise, which limits the movement of the affected joints. Lymphatic, neurotrophic, etc., so it is difficult to repair itself. Once the disease occurs, it often develops into osteoarthritis, and severe cases require artificial joint replacement. [0003] Treatment of this disease has previously been treated with a variety of therapeutic approaches, including debridement, bone marrow stimulation (eg, microfracture, subchondral bone drilling, etc.), autologous or allogeneic perichondrium and periosteum transplantation, osteochondral transplantation , articular car...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/24A61L27/50A61L27/54A61L27/58C12N5/0775
CPCA61L27/3834A61L27/24A61L27/3852A61L27/3895A61L27/50A61L27/54A61L27/58A61L2300/404A61L2300/412A61L2430/06C12N5/0663
Inventor 韦玉军李航陆宝石苏军吴远航
Owner 安徽安龙基因科技有限公司
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