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Preparation method for collagen membrane inoculated with autologous mesenchymal stem cells

A technology of bone marrow mesenchyme and collagen membrane, applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve the problems of poor treatment effect and high cost, and achieve simple operation, low cost, retention of curative effect and The effect of the operation

Inactive Publication Date: 2017-07-21
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

[0007] The purpose of the present invention is to solve the defects of poor therapeutic effect and high cost in the prior art, and provide a preparation method of collagen membrane inoculated with autologous bone marrow mesenchymal stem cells to solve the above problems

Method used

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preparation example Construction

[0036] The invention discloses a method for preparing a collagen membrane inoculated with autologous bone marrow mesenchymal stem cells. The specific steps are as follows:

[0037] (1) Under sterile conditions, draw out 9ml of bone marrow with a 20ml syringe containing 1ml of heparin sodium saline, remove the needle, add it to a culture bottle containing 10ml of DMEM high-glucose medium, and mix well to obtain a bone marrow suspension. spare;

[0038] (2) Under sterile conditions, take 100-200ml of peripheral blood from the patient, put it in a blood bag, seal it and bring it into the laboratory, then put the peripheral blood in a 50ml centrifuge tube, and centrifuge it at 3000rpm for 30min , take the supernatant, under the condition of 3000rpm, centrifuge the supernatant for 30min, remove the precipitate, filter and sterilize with a pore size of 0.22 μm, obtain autologous serum, and pack it for use;

[0039] (3) Slowly add the bone marrow suspension into the centrifuge tube ...

Embodiment 1

[0057] The invention discloses a method for preparing a collagen membrane inoculated with autologous bone marrow mesenchymal stem cells. The specific steps are as follows:

[0058] (1) Under sterile conditions, draw out 9ml of bone marrow with a 20ml syringe containing 1ml of heparin sodium saline, remove the needle, add it to a culture bottle containing 10ml of DMEM high-glucose medium, and mix well to obtain a bone marrow suspension. spare;

[0059] (2) Under sterile conditions, take 150ml of peripheral blood from the patient, put it in a blood bag, seal it and bring it to the laboratory, then put the peripheral blood in a 50ml centrifuge tube, centrifuge at 3000rpm for 30min, and take Supernatant, under the condition of 3000rpm, centrifuge the supernatant for 30min, remove the precipitate, filter and sterilize with 0.22 μm pore size, obtain autologous serum, and pack for use;

[0060] (3) Slowly add the bone marrow suspension into the centrifuge tube containing 20ml of cel...

Embodiment 2

[0077] The invention discloses a method for preparing a collagen membrane inoculated with autologous bone marrow mesenchymal stem cells. The specific steps are as follows:

[0078] (1) Under sterile conditions, draw out 9ml of bone marrow with a 20ml syringe containing 1ml of heparin sodium saline, remove the needle, add it to a culture bottle containing 10ml of DMEM high-glucose medium, and mix well to obtain a bone marrow suspension. spare;

[0079] (2) Under aseptic conditions, take 180ml of peripheral blood from the patient, put it in a blood bag, seal it and bring it to the laboratory, then put the peripheral blood in a 50ml centrifuge tube, centrifuge at 3000rpm for 30min, and take Supernatant, under the condition of 3000rpm, centrifuge the supernatant for 30min, remove the precipitate, filter and sterilize with 0.22 μm pore size, obtain autologous serum, and pack for use;

[0080] (3) Slowly add the bone marrow suspension into the centrifuge tube containing 20ml of cel...

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Abstract

The invention discloses a preparation method for a collagen membrane inoculated with autologous mesenchymal stem cells. The preparation method comprises the following concrete steps: carrying out uniform mixing so as to obtain a marrow suspension; preparing autologous serum; adding the marrow suspension into a centrifuge tube containing a cell separating medium and carrying out gradient centrifugation to obtain a sediment which is target cells; adding the target cells into 15 ml of a DMEM high-glucose medium containing FGF-2, the autologous serum and gentamycin, carrying out culturing in an incubator and replacing the DMEM high-glucose medium once every three days; after cell culture for 3 to 4 weeks, carrying out passage three times; subjecting cells to resuspension with normal saline; slowly adding the cell suspension onto the rough surface of a prepared pig-sourced I / III type collagen membrane with a plastic sleeve drop by drop; and allowing the collagen membrane to adsorb cells for 10 to 15 min so as to obtain the collagen membrane inoculated with the autologous mesenchymal stem cells. The collagen membrane inoculated with autologous mesenchymal stem cells prepared in the invention has the advantages of good treatment effect, convenience in treatment, low cost, etc.

Description

technical field [0001] The invention relates to the field of tissue engineering articular cartilage damage repair treatment, in particular to a method for preparing a collagen membrane inoculated with autologous bone marrow mesenchymal stem cells. Background technique [0002] Articular cartilage defects are relatively common in clinical practice. At present, the main methods for the treatment of articular cartilage defects have obvious defects. The current treatment methods for cartilage injuries include chondroplasty, bone marrow stimulation technology (ie, microfracture), autologous osteochondral transplantation, and autologous chondrocyte transplantation. therapy etc. [0003] Bone marrow stimulation technology is generally used clinically for smaller lesions in young patients, and it has also shown great success. Autologous osteochondral transplantation is usually used for patients with smaller lesions, and osteochondral transplantation is also used for larger cartilage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61L27/38A61L27/24A61L27/58
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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