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Preparation method of cell preparation for promoting regeneration and repair of traumas

A cell preparation and trauma technology, applied in the field of cell preparations, can solve the problems of lack of standardization and standardized approaches, etc.

Inactive Publication Date: 2016-12-07
ANHUI HUIEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some researchers have successfully isolated the culture and identification of mouse, rat, rabbit, and human tendon stem cells, but there is a lack of a standardized and standardized approach

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Step (1): Add 10% fetal bovine serum, 100U / mL penicillin, and 100U / mL streptomycin to DMEM medium to prepare a tendon stem cell culture solution;

[0031] Step (2), take the rat's hind limbs Achilles tendon, remove the tendon sheath and the part around the tendon with sterile scissors, wash the Achilles tendon tissue block with 0.1mol / L phosphate buffer solution for 3 times under aseptic conditions, and then wash the washed Achilles tendon Cut the tissue block to a volume of 0.5mm 3 ;

[0032] Step (3). Add the cut Achilles tendon tissue block in step (2) to the mixed solution of type I collagenase and 0.25% pancreatin solution. The volume ratio of the two solutions is 1:1. Suspend the Achilles tendon tissue. The solution was placed in a constant temperature water bath shaker at 37°C for 10 minutes, and the solid-liquid ratio was 1:5 to obtain the digested Achilles tendon tissue suspension;

[0033] Add 2 mL of the tendon stem cell culture solution configured in step (1) to t...

Embodiment 2

[0044] Step (1): Add 20% fetal bovine serum, 100U / mL penicillin, and 100U / mL streptomycin to DMEM medium to prepare tendon stem cell culture medium;

[0045] Step (2), take the rat's hind limbs Achilles tendon, use sterile scissors to remove the tendon sheath and the part around the tendon, and wash the Achilles tendon tissue block with 0.1mol / L phosphate buffer solution for 5 times under aseptic conditions. Cut the tissue block to a volume of 1mm 3 ;

[0046] Step (3). Add the cut Achilles tendon tissue block in step (2) to the mixed solution of type I collagenase and 0.25% pancreatin solution. The volume ratio of the two solutions is 1:1. Suspend the Achilles tendon tissue. The solution was placed in a constant temperature water bath shaker at 37°C for 30 minutes, and the solid-liquid ratio was 1:5 to obtain the digested Achilles tendon tissue suspension;

[0047] Add 2-5 mL of the tendon stem cell culture solution configured in step (1) to the digested Achilles tendon tissue susp...

Embodiment 3

[0058] Step (1): Add 15% fetal bovine serum, 100U / mL penicillin, and 100U / mL streptomycin to DMEM medium to prepare tendon stem cell culture medium;

[0059] Step (2), take the rat's hind limbs Achilles tendon, remove the tendon sheath and the part around the tendon with sterilized scissors, wash the Achilles tendon tissue block with 0.1mol / L phosphate buffer solution 4 times under aseptic conditions, and then wash the washed Achilles tendon Cut the tissue block to a volume of 0.75mm 3 ;

[0060] Step (3). Add the cut Achilles tendon tissue block in step (2) to the mixed solution of type I collagenase and 0.25% pancreatin solution. The volume ratio of the two solutions is 1:1. Suspend the Achilles tendon tissue. The solution was placed in a constant temperature water bath shaker at 37°C for 20 minutes and the solid-liquid ratio was 1:5 to obtain the digested Achilles tendon tissue suspension;

[0061] Add 4 mL of the tendon stem cell culture solution configured in step (1) to the di...

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Abstract

The invention discloses a preparation method of a cell preparation for promoting the regeneration and the repair of traumas. The preparation method comprises the steps of shearing and digesting heel tendon tissues, carrying out centrifuging to remove supernate, adding an obtained tendon stem cell population into a culture solution, carrying out resuspension and filtering, inoculating the culture solution into a culture plate for culturing, mixing the culture solution with platelet rich plasma, and adding normal saline, so as to prepare a tendon stem cell population. Tendon stem cells are separated and cultured, tendon stem cells cultured in vitro are mixed with the platelet rich plasma to prepare the cell preparation, the platelet rich plasma contains a large number of growth factors such as PDGF, EGF, TGF-beta1 and IGF which have great promotion effects to the repair of tendons, the platelet rich plasma is capable of promoting the propagation of tendon cells and the expression of collagens I and III in vivo, and the tendon cells and the collagens I and III are key components of the tendons, so that an experimental result has a certain guiding effect to the repair of tendon injury of a human body.

Description

Technical field [0001] The invention belongs to the technical field of cell preparations, and particularly relates to a preparation method of a cell preparation that promotes wound regeneration and repair. Background technique [0002] Tendinopathy is a chronic strain disease. Many studies have shown that tendinopathy caused by mechanical loading can cause a series of changes in tendon cells, such as changes in cell morphology, cell proliferation, apoptosis or necrosis, and increased local insulin-like growth factor (IGF-1) expression, among which apoptosis It is closely related to tendon degeneration. The abnormal increase in tendon apoptotic cells will affect the function of tendon cells, which in turn affects the synthesis and repair of collagen, resulting in loss of cell activity and reduced matrix synthesis. Protein synthesis disorders will weaken the tendon tissue and further increase the risk of rupture. [0003] In recent years, the development of stem cell therapy has op...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775A61K35/32A61K38/18A61K38/30A61P19/04A61L27/38A61L27/54
CPCC12N5/0662A61K35/32A61K38/1808A61K38/1841A61K38/1858A61K38/30A61L27/3804A61L27/386A61L27/3895A61L27/54A61L2300/252A61L2300/414A61L2430/10C12N5/0668C12N2500/84C12N2503/00C12N2509/00A61K2300/00
Inventor 张正亮
Owner ANHUI HUIEN BIOTECH
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