Method and device for preparing bone marrow mesenchymal stem cells-tube scaffold compound

A technology of bone marrow mesenchymal and pipeline scaffolds, which is applied in the field of preparation of bone marrow mesenchymal stem cells-pipe scaffold complexes, can solve the problems of poor pipeline histocompatibility, easy blockage of retrograde pipelines, time-consuming and other problems, and improve inoculation efficiency, Improves the effect of uniform distribution and amplification ability

Inactive Publication Date: 2011-09-21
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The former microsurgical technique is complex and time-consuming. Although it can reduce the occurrence of pipeline complications, pipeline complications can still reach 5% to 20%.
The biggest problem with the latter is that implanted

Method used

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  • Method and device for preparing bone marrow mesenchymal stem cells-tube scaffold compound
  • Method and device for preparing bone marrow mesenchymal stem cells-tube scaffold compound
  • Method and device for preparing bone marrow mesenchymal stem cells-tube scaffold compound

Examples

Experimental program
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Effect test

Embodiment 1

[0026] see figure 1 , figure 2 , the present invention constructs a perfusion culture device consisting of a peristaltic pump 1, a gas filter device 2, a liquid storage bottle 3 for containing a culture solution, a perfusion culture tank 4 and a silicone hose 5, and the silicone hose 5 is connected as a culture solution 6 perfusion channel various components. The peristaltic pump 1 adopts the 7524-55 type (Cole-Parmer Company, Shanghai) to generate perfusion power, the inner diameter of the silicone hose 5 (Cole-Parmer Company, Shanghai) is 0.35 cm, and the perfusion culture tank 4 is made of polypropylene compression molding , used to place the cell-pipe support complex 12, the inner diameter of the perfusion culture tank 4 is 8mm, the perfusion culture tank includes an upper cover 10 and a columnar tank body 11 that can be unloaded through the thread 14, and the top of the upper cover 10 is connected with a polypropylene tubular connection The mouth is the inlet of the cu...

Embodiment 2

[0031] A specific method adopted by the method of the present invention is: take isolated adult bone marrow and separate mononuclear cells, then perform purification and primary expansion culture of bone marrow mesenchymal stem cells, and prepare seed cells for tissue engineering based on bone marrow mesenchymal stem cells .

[0032]1. Take 3-5ml of isolated adult bone marrow, add 40ml of calcium and magnesium-free phosphate buffer solution (PBS), centrifuge at 900×g for 5 minutes, and discard the fat layer and supernatant. Precipitate with 8ml of α-MEM culture medium containing 10% (v / v) fetal bovine serum (FBS, Hangzhou Sijiqing Biotechnology Company, Hangzhou) to re-mix, slowly add to 4ml of Ficoll separation solution with a density of 1.077g / ml (Pharmacia Company, Shanghai), centrifuged at 900×g for 20 minutes, collected the mononuclear cell layer, washed 2-3 times with PBS, then resuspended the cells in α-MEM medium containing 10% fetal bovine serum and counted . Adjust...

Embodiment 3

[0035] Take the PCL pipe support that has been pre-treated with 95% alcohol for 2 hours, and then sterilized overnight with 75% alcohol. After blotted dry with sterile gauze, dry it with a slow-speed fan in an ultra-clean bench, and collect it for later use.

[0036] Take the bone marrow mesenchymal stem cells of the third generation above, and adjust the cell density to 2×10 with α-MEM culture medium containing 10% FBS. 6 After cells / ml, draw 400μl of cell suspension and evenly drop it on the outer surface of the outer wall of the pipeline support tube. into the outer wall of the pipe support. The inoculation method of uniform inoculation of 400 μl cell suspension plus vacuum treatment and the inoculation method of uniform inoculation of 400 μl cell suspension plus natural infiltration were used as controls.

[0037] After inoculation, take out the pipe support and place it in a six-well cell culture plate. After 2 hours, add 5ml of α-MEM containing 10% (v / v) fetal bovine se...

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Abstract

The invention provides a method and device for preparing a bone marrow mesenchymal stem cells-tube scaffold compound. In the invention, a cylinder in the inner center of a cylindrical groove blocks a central channel (inner hole) of a tube scaffold, so that culture fluid flowing through the culture fluid inlet on a top cap can completely flow across the outer wall pores of the tube scaffold, thus ensuring that cells in the outer wall pores of the tube scaffold acquire nourishment of the culture fluid. The method provided by the invention improves the inoculation efficiency of the bone marrow mesenchymal stem cells in the PCL (polycaprolactone) tube scaffold, and improves the uniform distribution and amplification capacities of the cells in the outer wall of the tube scaffold, a compound tube scaffold with bone marrow mesenchymal stem cells of even and high-density distribution can be obtained by the method, and the method is hopeful to be applied to repair of tube tissue damage clinically.

Description

(1) Technical field [0001] The invention relates to a method and a device for preparing a bone marrow mesenchymal stem cell-pipe scaffold compound. (2) Background technology [0002] Injury to duct tissue is a common clinical problem, such as duct leakage and duct stenosis in blood vessels or bile ducts. There are many ways to repair pipeline tissue damage: one is to improve surgical techniques, such as pipeline anastomosis under a surgical microscope to reduce anastomotic stenosis and pipeline leakage; the other is to implant stents in the pipeline. The former microsurgical technique is complex and time-consuming. Although it can reduce the occurrence of pipeline complications, pipeline complications can still reach 5% to 20%. The biggest problem with the latter is that implanted stents need to be taken out again through traumatic operations, and most of these stents are plastic or metal stents, which are poorly compatible with pipeline tissue and are prone to blockage and...

Claims

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Application Information

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IPC IPC(8): A61L27/38C12N5/0775C12M3/00
Inventor 王金福宗晨
Owner ZHEJIANG UNIV
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