Method for constructing fat stem cell membrane sheet and application

A technology of adipose stem cells and cell membranes, which is applied in the field of tissue engineering and regenerative medicine, to achieve the effective effect of repairing endometrial epithelium

Active Publication Date: 2018-09-04
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, how to effectively prevent the intrauterine adhesions formed after uterine injury, and enable the cell membrane to have a mechanism

Method used

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  • Method for constructing fat stem cell membrane sheet and application
  • Method for constructing fat stem cell membrane sheet and application
  • Method for constructing fat stem cell membrane sheet and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for constructing a fat stem cell membrane, comprising the steps of:

[0026] Step 1) selecting fat stem cells as seed cells to construct cell membranes;

[0027] Step 2) using endometrial cells for induced co-cultured cells;

[0028] Step 3) The adipose stem cell membrane is constructed by means of chemical induction and co-culture.

Embodiment 2

[0030] A method for constructing a fat stem cell membrane, comprising the steps of:

[0031] Step 1) Select adipose stem cells as seed cells to construct a cell membrane; said adipose stem cells: take 20ml of adipose tissue, wash it with PBS, cut it into pieces with ophthalmic scissors, add 20ml of 0.25% collagenase type 1 to digest on a constant temperature shaker at 37°C for 60min; Add an equal amount of α-mem complete medium containing 10% fetal bovine serum to stop the digestion, centrifuge at 1000 rpm for 5 minutes, remove the supernatant, add 10ml of α-mem complete medium containing 10% fetal bovine serum to resuspend and inoculate in a 75cm 2 In the culture flask, the medium was changed after 48 hours, and the adipose stem cells grew confluently and reached 80%-90% of the bottom area of ​​the flask, and were digested with trypsin for passage. The adipose stem cells are abbreviated as ADSCs.

[0032] Step 2) Endometrial cells are used to induce co-cultured cells; said e...

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Abstract

The invention relates to a fat stem cell membrane sheet, particularly relates to a method for constructing a fat stem cell membrane sheet and application, and belongs to the fields of tissue engineering and regenerative medicine. The method comprises the following steps: step 1) selecting and using fat stem cells as seed cells and constructing a cell membrane sheet; step 2) enabling endometrial cells to be applied to induced cocultured cells; and step 3) constructing the fat stem cell membrane sheet by adopting chemical induction and coculture modes. Exogenous factor (TGF-beta 1, EGF, PDGF-BBand 17-beta estradiol) are added and are compounded with the endometrial cells to perform coculture by utilizing a membrane sheet culture medium, human fat stem cells are induced to form membranes; the expression quantity of each epithelial marked mRNA of the membrane sheet which is induced by the membrane sheet culture system is up-regulated; and immunofluorescence shows that protein CK18 expresses positive and prompts that part of stem cells are transformed into epithelial cells. The membrane sheet induced by the system is more effective in the aspect of repairing endometrial epithelium.

Description

technical field [0001] The invention relates to an adipose stem cell membrane, in particular to a method and application for constructing an adipose stem cell membrane. It belongs to the field of tissue engineering and regenerative medicine. Background technique [0002] The normal endometrium is divided into basal layer and functional layer. The functional layer periodically sheds under the action of estrogen and progesterone, and is regenerated by the basal layer after shedding. Deposition, irreversible scar repair, resulting in intrauterine adhesions. [0003] 90% of the occurrence of intrauterine adhesions is related to the mechanical injury of the uterus during pregnancy or non-pregnancy, among which pregnancy-related uterine curettage is the main cause of intrauterine adhesions. According to the statistics of the World Health Organization (WHO), there are 40 million women in the world every year. Induced abortion, China's annual abortion reaches 14 million person-time...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/0775
CPCC12N5/0625C12N5/0667C12N2501/11C12N2501/135C12N2501/15C12N2506/1384
Inventor 王晓红肖西峰芦洁李博陈书强孙惠君杨芳王珺张琬琳王晶晶
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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