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Medicine for improving human skin, and preparation method and application of medicine

A drug and skin quality technology, applied in the field of medical cosmetology, can solve the problems of poor maintenance of conditioning effect, low success rate of cell planting, lack of cell growth microenvironment, etc., to improve skin condition, promote cell proliferation, and promote facial structure reconstruction Effect

Pending Publication Date: 2020-09-08
北京中瑞联合生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing technology or patent only uses a certain type of individual cells to regulate the face, because of the imbalance of the extracellular matrix of the face, the lack of a microenvironment for cell growth, and the low success rate of cell planting. Cell-to-cell interaction, conditioning effect, and maintenance time are poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Preparation of adipose-derived mesenchymal stem cells

[0028] 1) Collect adipose tissue into a 50mL centrifuge tube, wash with normal saline twice, and centrifuge to remove vascular tissue and red blood cells;

[0029] 2) Add an equal volume of preheated 0.1% type Ⅰ collagenase to the adipose tissue, rotate at 280r / min, and digest with shaking for 30min at 37°C;

[0030] 3) 300g centrifugal force to collect the bottom cell sediment;

[0031] 4) Resuspend cells in α-MEM medium containing 5% platelet supplement;

[0032] 5) After 36 hours, change the medium in full amount, remove non-adherent cells, add fresh complete medium, and change the medium in half the next day;

[0033] 6) When the cell confluency reaches 80-90%, digest and passage with 0.05% trypsin, continue to culture, and collect P3-P5 generation adipose-derived mesenchymal stem cells.

Embodiment 2

[0035] Skin fibroblast preparation

[0036] 1) Thoroughly disinfect behind the ear, and take 0.3cm×1cm skin tissue through local anesthesia;

[0037] 2) Wash the tissue with normal saline, and repeatedly knead the tissue with tweezers to remove the blood components inside the tissue;

[0038] 3) Pour 75% alcohol to disinfect for 30 seconds;

[0039] 4) The sterilized tissue was washed twice with normal saline, and the adipose tissue in the lower layer of the human skin was peeled off with tweezers to expose the dermis.

[0040] 5) Use a scalpel to cut human skin tissue into 0.3×0.5cm tissue pieces, paste them in 12-well plates, add 450uL α-MEM medium containing 5% platelet supplement, and change half of the medium every other day;

[0041] 6) When the cell confluency reaches 80-90%, digest and passage with 0.05% trypsin, continue to culture, and collect P3-P5 skin-derived fibroblasts.

Embodiment 3

[0043] Immune cell exosome preparation

[0044] 1) Take 30ml of venous blood, 1100g centrifugal force, increase the speed by 9 and decrease the speed by 9, and collect the serum;

[0045] 2) After the blood cell pellet was resuspended with an equal volume of PBS, it was added to an equal volume of 1.077 separation medium, and the centrifugal force was 900g, and the speed was increased by 7 and the speed was decreased by 7, to separate mononuclear cells;

[0046] 3) Collect buffy coat cells and wash them twice with PBS;

[0047] 4) Use DMEM / F12 containing autologous 3% plasma to resuspend the cells and adjust the cell density to 2X10 6 / mL;

[0048] 5) Add recombinant human CD3 antibody (final concentration 50ng / mL), human interleukin 2 (final concentration 1000 units / mL) and interferon (final concentration 1000 units / mL) to stimulate immune cell proliferation and exosomes secretion;

[0049] 6) Every other day, add fresh medium containing autologous 3% plasma, adjust the c...

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Abstract

The invention discloses a medicine for improving human skin, and a preparation method and application of the medicine. The medicine is composed of adipose tissue-derived stromal cells, skin fibroblasts, exosomes of peripheral blood immune cells and exosomes of adipose tissue-derived stromal cells, wherein the number of the adipose tissue-derived stromal cells is 5*10<6>-20*10<6> / mL, the number ofthe skin fibroblasts is 5*10<6>-20*10<6> / mL, and the protein concentration of the cell exosomes is 1-5 mg / mL. According to the medicine for improving skin quality of the human body, the fat exosomes are used for adjusting the face microenvironment; a good growth environment is provided for survival of the adipose tissue-derived stromal cells and fibroblasts; and the autologous adipose tissue-derived stromal cells are planted in combination with autologous skin fibroblasts, so that proliferation and differentiation of the skin cells can be promoted, the number of the face fibroblasts can be increased, secretion of collagen and fibrin can be increased, and comprehensive three-dimensional improvement and rejuvenation of the skin can be promoted.

Description

technical field [0001] The invention relates to the technical field of medical cosmetology, in particular to a medicine for improving human skin quality and its preparation method and application. Background technique [0002] Human skin has epidermis, dermis, and subcutaneous tissue. The skin covers the entire body surface and is the largest organ of the human body. It protects the body's ability to resist diseases. Fibroblasts in the dermis of the skin secrete a large amount of collagen and elastic fibers to keep the skin young. state. [0003] After the age of 25-30, due to sunlight, fatigue and other problems, the skin stem cells will decrease, and the aging process will begin to appear, especially the decrease of fibroblasts in human skin, resulting in weakened skin metabolism, pigmentation deposits and wrinkles. Protein synthesis is weakened, causing skin wrinkles, dryness, roughness and other problems, cell synthesis of elastic fibers is weakened, and the loss causes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/33A61K9/00C12N5/071C12N5/0775C12N5/078A61P17/00A61P17/16A61K35/28A61K35/14
CPCA61K35/28A61K35/33A61K35/14A61K9/0019C12N5/0667C12N5/0625C12N5/0634A61P17/00A61P17/16C12N2509/00C12N2509/10
Inventor 李志刚阮志黄宇唐雯
Owner 北京中瑞联合生物科技有限公司
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