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Application of mouse umbilical cord mesenchymal stem cells in protecting blood-brain barrier function after skin burn

A technology for mesenchymal stem cells and skin protection, applied in the field of immunology, can solve the problems of small amount of human mesenchymal stem cells and undisclosed blood-brain barrier treatment technology.

Active Publication Date: 2021-08-06
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the amount of human mesenchymal stem cells obtained is not large, and the prior art has not disclosed the treatment technology of umbilical cord mesenchymal stem cells for the damage of the blood-brain barrier after skin burns

Method used

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  • Application of mouse umbilical cord mesenchymal stem cells in protecting blood-brain barrier function after skin burn
  • Application of mouse umbilical cord mesenchymal stem cells in protecting blood-brain barrier function after skin burn
  • Application of mouse umbilical cord mesenchymal stem cells in protecting blood-brain barrier function after skin burn

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 In vitro isolation and culture of mouse umbilical cord mesenchymal stem cells

[0029] The source of stem cells is obtained by in vitro expansion and culture of fetal mouse-derived cells

[0030] (1) Obtain the umbilical cord of the fetal mouse;

[0031] (1.1) Execute the C57 pregnant mice with a gestational age of 19-20 days, and soak them in alcohol for disinfection;

[0032] (1.2) Cut open the abdomen of the mouse to expose the abdominal cavity, separate the fetal mouse and place it in a sterile petri dish or operation tray;

[0033] (1.3) Cut the fetal membranes to expose the fetal mouse and placenta, expose the umbilical cord, fix the two ends of the umbilical cord with 8cm hemostats, cut the umbilical cord outside, and put the separated umbilical cord into a sterile culture dish, repeat the above steps until all The fetal mouse umbilical cord was separated;

[0034] (2) Put the umbilical cord of the fetal mouse into a petri dish, cut it into pieces, p...

Embodiment 2

[0058] Example 2 Preparation of mouse umbilical cord mesenchymal stem cell drug for treating blood-brain barrier permeability after scalding in mice

[0059] 1) Count the freshly obtained P3 mouse cells or the thawed P3 mouse cells using a cell counter, about 5-6*10 6 / ml;

[0060] 2) Wash 2 times with 2ml PBS, add 1ml trypsin, and incubate in a 38°C incubator for 2 minutes

[0061] 3) After observing all the suspension of cells under a microscope, add 3ml DMEM (containing 10% fetal bovine serum and 1% penicillin) and mix well, then inhale into a 15ml centrifuge tube;

[0062] 4) Put it into a centrifuge for centrifugation, the centrifugation speed is 1000rpm, and the centrifugation time is 5min;

[0063] 5) Take out the centrifuge tube, remove the supernatant, add 500ul of normal saline, and mix well to obtain an injection for protecting the damaged blood-brain barrier after skin burns.

Embodiment 3

[0064] Example 3 Application of Mouse Umbilical Cord Mesenchymal Stem Cells to Treat Blood Brain Barrier Permeability Changes in Scalded Mice

[0065] 1. Establishment of scald model in mice

[0066] Healthy 7-week-old mice were selected and randomly divided into 6 groups, 6 mice in each group. The mouse was anesthetized and the back skin was removed, and the skin hair on the back of the mouse was removed with a shaver, the size of which was 2 cm in diameter. Then, a constant temperature and constant pressure scalding apparatus was used to scald the mouse's back for 3 seconds at a scalding temperature of 90-94° C., with a scalding pressure of 500 g.

[0067] 2. Select one group as the control group, inject 10kDa fluorescent tracer into each mouse at 0.5ml / kg through the rat tail vein before scalding in the control group, and inject 10kDa fluorescent tracer into each mouse at 0, 1, 3, 6, 12 hours after scalding After femoral artery bloodletting and killing, the brain tissue o...

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Abstract

The invention relates to the application of mouse umbilical cord mesenchymal stem cells in the preparation of a drug for protecting the damaged blood-brain barrier after skin burns; and a method for preparing the drug for protecting the damaged blood-brain barrier after skin burns. Experiments have proved that mouse umbilical cord mesenchymal stem cells can effectively improve the permeability of the blood-brain barrier after scalding in mice.

Description

technical field [0001] The invention relates to the field of immunology, in particular to the application of mouse umbilical cord mesenchymal stem cells in protecting blood-brain barrier function damage after skin scalding, especially the application of protecting blood-brain barrier function after skin scalding in mice. Background technique [0002] Mesenchymal stem cells originally referred to bone marrow stromal cells. Later studies found that stem cells with similar biological characteristics and the same immune phenotype as bone marrow stromal cells were extracted from fetal liver, lung, heart and other solid organs, umbilical cord and cord blood. Therefore, cells derived from mesenchymal tissue that have similar biological properties to bone marrow MSCs and have multilineage differentiation potential are collectively referred to as mesenchymal stem cells. MSCs are multipotent adult stromal cells that can differentiate into a variety of tissues of mesenchymal origin, su...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775
CPCA61K35/51A61P17/02A61P25/00C12N5/0668
Inventor 杨思明杨劼刘煜凡马奎付小兵
Owner GENERAL HOSPITAL OF PLA
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