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Method for culturing stem cell factors for beauty

A stem cell factor and culture method technology, applied in animal cells, culture process, tissue culture and other directions, can solve the problems of stem cell viability, low differentiation efficiency, low cytokine production, etc., to facilitate product separation, improve proliferation efficiency, reduce effect of loss

Active Publication Date: 2021-06-15
张若冰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the stem cells cultivated by the current method have low viability and differentiation efficiency, and the production of cytokines is low.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A method for cultivating stem cell factors for beauty treatment, comprising the following steps:

[0031] (1) The tissue blocks containing mesenchymal stem cells were placed in culture flasks, and then serum-free medium was added, at 37°C, 5% CO 2 The incubator can only be cultivated for 7 days;

[0032] (2) When the cell confluency reaches 70%, discard the serum-free medium, wash the bottle wall with normal saline, then add 0.25% trypsin to evenly infiltrate the cell adherent surface, incubate at room temperature for 5 minutes, shake quickly and blow after the cells become round Cell adherent surface, obtain cell suspension, centrifuge to discard supernatant, and then use 6000 cells / cm 2 The cell density continued to be subcultured until the cells were subcultured to the 4th generation;

[0033] (3) When the confluence of the cells obtained in step (2) reaches 80%, replace with the induction medium and continue at 37°C, 5% CO 2 Cultivate in the environment for 72 ho...

Embodiment 2

[0037] A method for cultivating stem cell factors for beauty treatment, comprising the following steps:

[0038] (1) The tissue blocks containing mesenchymal stem cells were placed in culture flasks, and then serum-free medium was added, at 37°C, 5% CO 2 The incubator can only be cultivated for 7 days;

[0039] (2) When the cell confluence reaches 80%, discard the serum-free medium, wash the bottle wall with normal saline, then add 0.25% trypsin to evenly infiltrate the adherent surface of the cells, incubate at room temperature for 5 minutes, shake quickly and pipette after the cells become round Cell adherent surface, obtain cell suspension, centrifuge to discard supernatant, and then use 6000 cells / cm 2 The cell density continued to be subcultured until the cells were subcultured to the 4th generation;

[0040] (3) When the confluence of the cells obtained in step (2) reaches 85%, replace with the induction medium and continue at 37°C, 5% CO 2 Cultivate in the environmen...

Embodiment 3

[0044]A method for cultivating stem cell factors for beauty treatment, comprising the following steps:

[0045] (1) The tissue blocks containing mesenchymal stem cells were placed in culture flasks, and then serum-free medium was added, at 37°C, 5% CO 2 The incubator can only be cultivated for 7 days;

[0046] (2) When the cell confluence reaches 80%, discard the serum-free medium, wash the bottle wall with normal saline, then add 0.25% trypsin to evenly infiltrate the adherent surface of the cells, incubate at room temperature for 5-10 minutes, shake rapidly after the cells become round And blow the cell adherent surface to get the cell suspension, centrifuge to discard the supernatant, and then use 6000 cells / cm 2 The cell density continued to be subcultured until the cells were subcultured to the 4th generation;

[0047] (3) When the confluence of the cells obtained in step (2) reaches 85%, replace with the induction medium and continue at 37°C, 5% CO 2 Cultivate in the ...

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Abstract

The invention discloses a method for culturing stem cell factors for beauty. The method comprises the following steps of (1) placing mesenchymal stem cells in a serum-free medium, sub-culturing to 4 generations, flushing with normal saline, then adding digestive juice, and carrying out digestion counting; (2) when the fusion degree of the cells obtained in the step (1) reaches 80-90%, replacing with an induction culture medium, and continuing to culture; (3) filtering the induction culture medium obtained in the step (2) for 3-4 times, merging filtrates, and concentrating to 1 / 10-1 / 5 of the original volume; adding normal saline containing sodium citrate to complement to the original volume, filtering again, collecting filtrate, and concentrating the filtrate to 1 / 20-1 / 10 of the original volume; and (4) filtering the product obtained in the step (3) by using a filter membrane with the molecular weight cutoff of 8-14KD, and collecting to obtain the stem cell factor concentrated solution. According to the method, the content of cell factors in the prepared concentrated solution can be effectively increased.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a method for cultivating stem cell factors for beauty treatment. Background technique [0002] In recent years, with the deepening of research on mesenchymal stem cells, its secreted factors have become a hot spot for researchers. Mesenchymal stem cells can secrete a variety of biologically active cytokines, which can effectively regulate body cell signal transduction, activate human stem cells, and then physiologically repair or replace damaged, diseased and aging cells in the body. [0003] The application of stem cells in cosmetology is the earliest use of stem cells in plastic surgery. It is a stem cell cosmetology technology that has been studied more at present. The types of stem cells that are widely used in plastic surgery are mainly human umbilical cord mesenchymal stem cells, human adipose-derived mesenchymal stem cells, etc. [0004] However, the mec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/64A61K8/99A61K8/34A61K8/60A61K8/63A61Q19/00C12N5/0775
CPCA61K8/64A61K8/99A61K8/60A61K8/345A61K8/63C12N5/0662A61Q19/00C12N2500/02C12N2500/30C12N2500/32A61K2800/84A61K2800/805
Inventor 张若冰谢惠明张茜玥谢家琪张杰韩飞徐根艳方民
Owner 张若冰
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