Application of stem cell conditioned medium in preparation of medicine for treating inflammatory skin

A conditioned medium and technology for inflammatory skin diseases, applied in the field of biomedicine, can solve the problems of poor quality inflammatory skin effects, etc., and achieve significant technological progress, inhibit lesion, and inhibit skin inflammation

Pending Publication Date: 2021-12-24
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the above-mentioned technical problems in the prior art, the present invention provides the application of the stem cell regulated medium in the preparation of medicines for treating inflammatory skin, which will solve the problem of the quality of the medicines in the prior art for the treatment of inflammatory skin Poor skin effect due to technical issues

Method used

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  • Application of stem cell conditioned medium in preparation of medicine for treating inflammatory skin
  • Application of stem cell conditioned medium in preparation of medicine for treating inflammatory skin
  • Application of stem cell conditioned medium in preparation of medicine for treating inflammatory skin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Preparation of human umbilical cord mesenchymal stem cell conditioned medium for skin medicine (UMSCM)

[0035] 1) Isolation and culture of human umbilical cord mesenchymal stem cells: After cleaning the human umbilical cord with sterile saline, remove the umbilical artery and umbilical vein blood vessels with tooth forceps, and tear off the remaining tissue after the epidermis, cut into Tissue blocks with a size of 3mm, and then, adopt the method of tissue climbing, let the tissue stick in the culture dish, add the medium alpha-MEM containing the platelet lysate with a concentration of 5% by volume. The medium was changed slowly after 7 days, and the cells crawled out of the tissue block after about 2 weeks. Subculture was carried out when the cells reached 90% density, and the morphology of umbilical cord mesenchymal stem cells was shown in figure 1 .

[0036] 2) Collect stem cell conditioned medium: continue to culture umbilical cord mesenchymal stem cel...

Embodiment 2

[0038] Example 2: Preparation of human amnion epithelial cell conditioned medium for skin medication (AECM)

[0039] 1) Isolation and culture of amniotic epithelial stem cells: Isolate the amniotic membrane from human placenta tissue, wash it with sterile saline, separate the epithelial layer, the layer close to the fetus, and then digest it with 0.25% trypsin for 30 Minutes, and then digested with 0.1g / L collagenase for 1 hour, finally, filtered and centrifuged to obtain primary amniotic epithelial stem cells, in DMEM / F12 medium containing 5% platelet lysate by volume, when cells were found Colony growth, when the adherent cells reached 80%-90% confluence, they were digested with 0.25% trypsin in mass percentage concentration, and then subcultured; the morphology characteristics of amniotic membrane epithelial stem cells were as follows figure 1 shown.

[0040] 2) Collect stem cell conditioned medium: continue to culture umbilical cord mesenchymal stem cells or amniotic memb...

Embodiment 3

[0042] Example 3: Quality identification of human umbilical cord mesenchymal stem cells and human amniotic epithelial cell conditioned medium

[0043] 1) The stem cell conditioned medium is subjected to sterility testing, detection of mycoplasma, endotoxin, etc.

[0044] 2) The stem cell conditioned medium was subjected to independent and allergy testing experiments.

[0045] 3) BCA quantification is performed on the stem cell conditioned medium, and the total protein amount and protein quantification of important factors in it are determined. Specifically, the content detection of active ingredients IL-1ra, IL-10, KGF, bFGF, for IL-1ra content is 400-1000pg / ml, IL-10 is 50-200pg / ml, KGF, EGF, bFGF and content are 50-200pg / ml.

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Abstract

The invention provides application of a stem cell conditioned medium in preparation of a medicine for treating inflammatory skin. The preparation method of stem cells comprises the following steps of taking amnion and umbilical cord tissues in human placenta tissues as raw materials, and then obtaining amniotic epithelial cells and umbilical cord mesenchymal stem cells through methods such as cutting, digestion and the like; then, inoculating in a culture medium containing a platelet lysate; carrying out passage on the obtained primary cells to the third generation, and discarding the culture medium containing the platelet lysate when the cell confluence degree reaches 90%; replacing the culture medium with a serum-free basic culture medium, and continuously culturing for 20-36 hours; and collecting the conditioned culture solution in which the cells are cultured, and centrifuging to remove the cells and cell debris, so as to obtain the amniotic epithelial cell conditioned medium (AECM) and umbilical cord mesenchymal stem cell conditioned culture medium (UMSCM). The AECM and the UMSCM can inhibit inflammatory cells of the skin of a patient from infiltrating the skin, so that epidermal hyperplasia and inflammatory response are inhibited.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a medicine for treating psoriasis, in particular to the use of a stem cell conditioned medium in preparing medicine for treating inflammatory skin. Background technique [0002] Inflammatory skin diseases are a series of recurrent skin diseases caused by the immune system's stimulation of environmental factors and abnormal activation of the immune system. It has become one of the major health problems in the world, and the proportion of healthcare expenditure on the treatment of such diseases increase year by year. Psoriasis, a typical refractory inflammatory skin disease, is a common chronic inflammatory skin disease characterized by massive infiltration of various types of immune cells dominated by T lymphocytes and abnormal epidermal keratinocytes Prolongation of epidermal spines caused by hyperplasia, incomplete differentiation of keratinocytes caused by abnormal differentiation, et...

Claims

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Application Information

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IPC IPC(8): A61K35/50A61K35/28A61P17/00A61P29/00A61P17/06A61P25/00A61P17/10A61P17/08A61P37/08C12N5/0775C12N5/071C12N5/073
CPCA61K35/50A61K35/28A61K9/0014A61P17/00A61P29/00A61P17/06A61P25/00A61P17/10A61P17/08A61P37/08C12N5/0668C12N5/0625C12N5/0605C12N2509/00C12N2509/10C12N2501/2313C12N2501/2311C12N2501/231C12N2501/117C12N2501/115C12N2501/2301C12N2501/15C12N2501/21C12N2501/11C12N2501/135C12N2501/58
Inventor 徐辉明杨孟波高维强王岚琦鞠强
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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