Preparation method and application of collagen scaffold composite bone marrow-derived mesenchymal stem cells (BMSCs)

A technology of bone marrow mesenchymal and collagen scaffolds, applied in the field of obstetrics and gynecology, can solve the problems of inability to promote the growth of scar intima, affect the curative effect, and difficult to locate, so as to increase the thickness of intima and local blood vessel density, and promote scarred uterus Intimal repair, good biocompatibility effect

Active Publication Date: 2014-04-09
YANTAI ZHENGHAI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, severe damage to the endometrium cannot solve the problem of endometrial scarring, cannot achieve functional repair of the endometrium, and is very prone to re-adhesion
[0005] 2. Recently, a small sample study reported that after the separation of intrauterine adhesions, human fresh amniotic membrane was used to cover the wound. This method cannot induce the functional repair of the scar endometrium itself, and the amniotic membrane still has risks such as immune inflammation and induced infecti

Method used

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  • Preparation method and application of collagen scaffold composite bone marrow-derived mesenchymal stem cells (BMSCs)
  • Preparation method and application of collagen scaffold composite bone marrow-derived mesenchymal stem cells (BMSCs)
  • Preparation method and application of collagen scaffold composite bone marrow-derived mesenchymal stem cells (BMSCs)

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Composite culture of BMSCs and collagen scaffolds

[0037] 1. Equipment, materials and reagents used

[0038] Ultra clean bench, CO 2 Incubator, inverted microscope, centrifuge, scanning electron microscope, critical point dryer, tissue embedding freezer, paraffin slicer, automatic tissue staining machine, cell culture bottle, 6-well plate, centrifuge tube, L-DMEM culture medium, Fetal bovine serum, penicillin / streptomycin, trypsin, formaldehyde, glutaraldehyde, xylene, concentration gradient ethanol, hematoxylin, eosin, etc.

[0039] 2. BMSCs maintain cell viability

[0040] (1) When cultured until 90% of the adherent cells are confluent, subculture is carried out;

[0041] (2) Remove the old culture medium, add PBS to wash away the residual culture medium, and remove the PBS;

[0042] (3) Add 1ml of trypsin and shake the flask gently;

[0043] (4) Observing under an inverted microscope, it was found that the cytoplasm retracted and the intercellular space increas...

Embodiment 2

[0062] Collagen Scaffold Composite BMSCs Used in the Repair Treatment of Severe Uterine Injury Animal Model

[0063] 1. Identification of BMSCs before use

[0064] (1) Sterility test: If there is no abnormality during the culture process, 3 days before the compounding of BMSCs and collagen scaffolds, after the last liquid change, a sterility test is performed on the supernatant in each culture bottle to culture bacteria and mold.

[0065] (2) Appearance and observation under the microscope: on the day when BMSCs and collagen scaffolds are combined, if the sterility test reports no bacteria and mold growth, observe the color of the supernatant in the culture bottle, and observe the cell morphology and cell morphology under an inverted microscope. Density, presence or absence of bacterial or mold growth. If there is any doubt, another sample must be taken for sterility test to cultivate bacteria and mold.

[0066] (3) Proportion of live cells: On the day when BMSCs and collag...

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Abstract

The invention relates to a preparation method and application of collagen scaffold composite bone marrow-derived mesenchymal stem cells (BMSCs). The preparation method comprises the steps of preparing single cell suspension after trypsinizing the BMSCs, uniformly dropwise adding the single cell suspension to a collagen scaffold, putting the collagen scaffold into an incubator to be cultured, and adding an L-DMEM complete medium to continue culture, thus obtaining the collagen scaffold composite BMSCs. The preparation method has the advantages that the following defects are overcome: endometria are seriously injured as intrauterine adhesion is mechanically separated by adopting hysteroscopic surgery, intrauterine devices or anti-adhesion materials are put after the surgery and estrogens are given after the surgery to promote intima growth; the problem of intima scars can not be solved; functional intima repair can not be achieved; adhesion is very easy to happen again. As active ingredients for treating serious endometrium injury, the BMSCs are convenient to obtain, secrete growth factors to improve the local microenvironment and immunoregulation, have good biocompatibility, degradability and safety, promote scarred endometrium repair and increase the intima thickness and local blood vessel density.

Description

technical field [0001] The invention belongs to the field of obstetrics and gynecology, and relates to a preparation method and application of collagen scaffold composite bone marrow mesenchymal stem cells. Background technique [0002] When the endometrium is severely damaged, the functional repair of the endometrium is obstructed, replaced by connective tissue hyperplasia and repair, fibrosis of the anterior and posterior walls of the uterus, and scar formation. Clinically, the main manifestations are intrauterine adhesions, amenorrhea, and uterine infertility. There is no effective treatment. [0003] Prior to the present invention, the treatment for severe endometrial injury was as follows: [0004] 1. The common clinical method is to mechanically separate intrauterine adhesions through hysteroscopic surgery, place intrauterine devices or anti-adhesion materials (mainly hyaluronic acid, Intercoat gel) after surgery, and give estrogen after surgery to promote intrauterin...

Claims

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Application Information

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IPC IPC(8): A61L31/04A61L31/16
Inventor 胡娅莉戴建武李新安丁利军
Owner YANTAI ZHENGHAI BIO TECH
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