Preparation method and application of cellular composite endometrial repair gel

A technology for endometrial repair and endometrium, applied in the field of biomedicine, can solve problems such as re-adhesion, poor clinical treatment effect, and insufficient time for cell repair

Pending Publication Date: 2020-12-08
山西宾大干细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the treatment methods for intrauterine adhesions are generally not effective in clinical treatment, and the

Method used

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Experimental program
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Effect test

specific Embodiment 1

[0108]A method for preparing cell composite endometrial repair gel, including the following steps:

[0109]Step 1: Extract and culture endometrial stem cells to obtain endometrial stem cells;

[0110]Step 2: Extraction and culture of mesenchymal stem cells to obtain mesenchymal stem cells;

[0111]Step 3. Preparation of temperature-sensitive chitosan hydrogel;

[0112]Step 4. Preparation of cell complex endometrial repair gel.

[0113]Further, step one includes the following steps:

[0114](1) Under aseptic conditions, take the endometrial tissue and use Ca-free2+And Mg2+Rinse at least three times with PBS buffer, then transfer to a sterile petri dish and cut it to a volume less than 1mm3Small piece

[0115](2) Add 2 times the volume of 0.1v / v% collagenase aqueous solution to the sterile petri dish of step (1), shake and digest in a water bath at 37°C for 50 min, collect the digestion solution in a centrifuge tube, and transfer it to the centrifuge tube Add DMEM / F12 complete medium to stop the digestion...

specific Embodiment 2

[0180]It is roughly the same as the specific embodiment 1, except that the industrial control conditions in step two are different:

[0181]The mesenchymal stem cells in step two are umbilical cord mesenchymal stem cells, and step two includes the following steps:

[0182](1) Under aseptic conditions, take a healthy umbilical cord specimen of 10 cm and place it in PBS buffer containing 1v / v% double antibody;

[0183](2). Move the umbilical cord specimen obtained in step (1) to the ultra-clean workbench, and rinse the umbilical cord specimen with PBS buffer repeatedly to remove residual blood;

[0184](3) After peeling off the outer membrane of the umbilical cord specimen and umbilical cord arteries and veins in a petri dish, cut the remaining tissue block to a volume less than 1mm3;

[0185](4) Add DMEM / F12 complete medium to the petri dish, place it at 37℃, 5v / v% CO2, Cultivate in a carbon dioxide incubator with saturated humidity, and perform a half volume exchange after 4 days. After the cells ...

specific Embodiment 3

[0189]It is roughly the same as the specific embodiment 1, except that the industrial control conditions in step two are different:

[0190]The mesenchymal stem cells in step two are umbilical cord mesenchymal stem cells, and step two includes the following steps:

[0191](1) Under aseptic conditions, take a 20cm healthy umbilical cord specimen and place it in PBS buffer containing 1v / v% double antibody;

[0192](2). Move the umbilical cord specimen obtained in step (1) to the ultra-clean workbench, and rinse the umbilical cord specimen with PBS buffer repeatedly to remove residual blood;

[0193](3) After peeling off the outer membrane of the umbilical cord specimen and umbilical cord arteries and veins in a petri dish, cut the remaining tissue block to a volume less than 1mm3;

[0194](4) Add DMEM / F12 complete medium to the petri dish, place it at 37℃, 5v / v% CO2, Cultivate in a carbon dioxide incubator with saturated humidity. After 4 days, perform a half volume change. After the cells are found...

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Abstract

The invention discloses a preparation method of a cellular composite endometrial repair gel. The method comprises the following steps of 1. extracting and culturing endometrial stem cells to obtain endometrial stem cells; 2, extracting and culturing mesenchymal stem cells to obtain the mesenchymal stem cells; 3, preparing temperature-sensitive chitosan hydrogel; and 4, preparing the cellular composite endometrial repair gel. The invention also discloses application of the cellular composite endometrial repair gel in treating intrauterine adhesion. The invention has the following advantages that 1, autologous stem cell transplantation basically eliminates the risk of rejection; 2, the autologous endometrial stem cells and the mesenchymal stem cells are subjected to composite transplantation, so that regeneration of endometrium, microvessels and mesenchyme is accelerated, and the endometrium repair time is obviously shortened; and 3, the temperature-sensitive gel can be formed after local injection, which can only only provide a three-dimensional retention space for the stem cells, but also prevent the adhesion of regenerated mucous membranes.

Description

Technical field[0001]The invention relates to the field of biomedicine, in particular to a preparation method and application of a cell composite endometrial repair gel.Background technique[0002]According to literature reports, the incidence of intrauterine adhesions caused by artificial abortion and curettage is as high as 25%-30%, and the incidence of postoperative re-adhesion is very high, especially for severe intrauterine adhesions, and the postoperative re-adhesion rate is as high as 62.5% .[0003]How to completely cure intrauterine adhesions is a difficult clinical problem. In recent years, scholars at home and abroad have proposed that the occurrence of intrauterine adhesions is related to the damage of endometrial stem cells. Endometrial injury leads to the reduction of endometrial stem cells or impaired function. The endometrial is unable to regenerate completely, leading to endometrial scar repair and ultimately to the uterus. Occurrence of cavity adhesions.[0004]As a kind...

Claims

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Application Information

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IPC IPC(8): A61K35/48A61K9/06A61K47/36A61K47/24A61P41/00A61P15/00C12N5/0775C12N5/074A61K35/28
CPCA61K35/28A61K35/48A61K9/06A61K47/36A61K47/24A61K9/0034A61P41/00A61P15/00C12N5/0682C12N5/0668C12N5/0667C12N5/0663C12N2509/00A61K2300/00
Inventor 傅松涛
Owner 山西宾大干细胞生物科技有限公司
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