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Method for three-dimensionally culturing bone marrow mesenchymal stem cells based on Collagel scaffold method

A bone marrow mesenchymal, three-dimensional culture technology, applied in the field of biotechnology and cell culture, can solve the problems of unsatisfactory three-dimensional cell culture technology, and achieve the effect of ensuring tissue moisture, enhancing adhesion ability, and not easy to fall off

Inactive Publication Date: 2020-09-18
SHANDONG MEDICAL BIO TECH RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] For this reason, the technical problem to be solved by the present invention is to provide a method for three-dimensional culture of bone marrow mesenchymal stem cells based on the Collagel gel scaffold method, so as to solve the problem of unsatisfactory effect of traditional three-dimensional cell culture technology in the prior art

Method used

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  • Method for three-dimensionally culturing bone marrow mesenchymal stem cells based on Collagel scaffold method
  • Method for three-dimensionally culturing bone marrow mesenchymal stem cells based on Collagel scaffold method
  • Method for three-dimensionally culturing bone marrow mesenchymal stem cells based on Collagel scaffold method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The Collagel culture medium matrix described in the present embodiment comprises following composition:

[0044] Reagent A: 5×MEM, 288 μl;

[0045] Reagent B: FBS, 144 μl;

[0046] Reagent C: Hepes (1M), 36 μl;

[0047] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;

[0048] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.

[0049] Take 288 μl of Reagent A (5×MEM), 144 μl of Reagent B (FBS) and 36 μl of Reagent C (Hepes, 1M) in Test Tube 1 for thorough mixing and set aside; another 1260 μl of Collagel (3mg / ml COL1 with 0.01M HCl mixed solution) and 72 μl of reagent D (0.1M NaOH and 5×MEM mixed solution) were placed in test tube 2 for thorough mixing, and the pH of the mixed solution was adjusted to 7.0 for later use; the mixed solutions in test tube 1 and test tube 2 were mixed to obtain Required Collagel culture medium, spare. It should be noted that the storage and operation of all the above reagents need to be kept at 4°C or on ice.

[0050] Select bone marro...

Embodiment 2

[0055] The Collagel culture medium matrix described in the present embodiment comprises following composition:

[0056] Reagent A: 5×MEM, 288 μl;

[0057] Reagent B: FBS, 144 μl;

[0058] Reagent C: Hepes (1M), 36 μl;

[0059] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;

[0060] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.

[0061] Take 288 μl of Reagent A (5×MEM), 144 μl of Reagent B (FBS) and 36 μl of Reagent C (Hepes, 1M) in Test Tube 1 for thorough mixing and set aside; another 1260 μl of Collagel (3mg / ml COL1 with 0.01M HCl mixed solution) and 72 μl reagent D (0.1M NaOH and 5×MEM mixed solution) were placed in test tube 2 for thorough mixing, and the pH of the mixed solution was adjusted to 7.0-7.2 for later use; mix the mixed solutions in test tube 1 and test tube 2 , to obtain the required Collagel culture substrate, and set aside. It should be noted that the storage and operation of all the above reagents need to be kept at 4°C or on ice.

[0062] Select...

Embodiment 3

[0066] The Collagel culture medium matrix described in the present embodiment comprises following composition:

[0067] Reagent A: 5×MEM, 288 μl;

[0068] Reagent B: FBS, 144 μl;

[0069] Reagent C: Hepes (1M), 36 μl;

[0070] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;

[0071] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.

[0072] Take 288 μl of Reagent A (5×MEM), 144 μl of Reagent B (FBS) and 36 μl of Reagent C (Hepes, 1M) in Test Tube 1 for thorough mixing and set aside; another 1260 μl of Collagel (3mg / ml COL1 with 0.01M HCl mixed solution) and 72 μl reagent D (0.1M NaOH and 5×MEM mixed solution) were placed in test tube 2 for thorough mixing, and the pH of the mixed solution was adjusted to 7.0-7.2 for later use; mix the mixed solutions in test tube 1 and test tube 2 , to obtain the required Collagel culture substrate, and set aside. It should be noted that the storage and operation of all the above reagents need to be kept at 4°C or on ice.

[0073] Select...

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Abstract

The invention belongs to the technical field of biotechnology and cell culture, and particularly relates to a method for three-dimensionally culturing bone marrow mesenchymal stem cells based on a Collagel scaffold method. According to the method disclosed by the invention, a Collagel scaffold is used as a three-dimensional matrix, and the bone marrow mesenchymal stem cells and the Collagel are mixed to form cell suspension for culturing, so that a tissue-like physical and spatial three-dimensional structure required by cell growth can be better simulated; the microenvironment of cell growth under different biomechanical effects of an in-vivo physiological environment can be better simulated, the biological functions of growth, proliferation, adhesion, differentiation and the like of the bone marrow mesenchymal stem cells are promoted, and the method is more suitable for researches in the fields of medical tissue engineering and biomechanics, such as intervertebral disc tissue repair and fibrous ring tissue regeneration.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and cell culture, and in particular relates to a method for three-dimensionally culturing bone marrow mesenchymal stem cells based on a Collagel gel scaffolding method. Background technique [0002] Bone marrow mesenchymal stem cells are bone marrow stromal stem cells, which have strong self-renewal and differentiation potential. Potential cell subsets. BMSCs have a comprehensive secretory function, can secrete growth factors, chemokines, adhesion molecules, extracellular matrix molecules, etc., and through the interaction of these factors, regulate the bone marrow and blood microenvironment, guide stem cells to migrate to damaged tissues, and proliferate , differentiation, and promote the repair and regeneration of damaged tissues. Based on these biological characteristics, BMSCs have become ideal seed cells for repairing damaged tissues in tissue engineering. [0003] Studies have shown...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12N5/02
CPCC12N5/0663C12N2513/00C12N2533/54C12N2533/50C12N2533/52C12N2501/999
Inventor 王丹丹师彬刘凡杰曹盛楠孙士飞
Owner SHANDONG MEDICAL BIO TECH RES CENT
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