Method for rapidly increasing exosome yield of adipose tissue-derived stromal cells

A technology of stem cells and exosomes, applied in the field of biomedicine, can solve the problems of few stem cells, limited application, and difficulty in self-repair and clinical treatment.

Pending Publication Date: 2021-03-26
重庆市铂而斐细胞生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the amount of stem cells is very small, and with the increase of individual age, the number of stem cells and the secretion activity gradually decrease, resulti...

Method used

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  • Method for rapidly increasing exosome yield of adipose tissue-derived stromal cells
  • Method for rapidly increasing exosome yield of adipose tissue-derived stromal cells
  • Method for rapidly increasing exosome yield of adipose tissue-derived stromal cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] A method to rapidly increase the production of exosomes from adipose-derived mesenchymal stem cells:

[0060] 1. Prepare pre-activation solution: 0.8% laminin, 0.1% collagen, 0.6% fibronectin, 98.5% solvent, mix well and set aside.

[0061] 2. Take the fat after liposuction to separate and cultivate adipose-derived mesenchymal stem cells; take the P3-P5 generation of adipose-derived mesenchymal stem cells, count the cells and inoculate them in a culture flask at an equal density. The medium was Lonza serum-free medium + PALL's serum substitute + Gibco's glutamine mixture.

[0062] 3. Add 1 ng / mL pre-activation solution to the culture bottle, and treat the adipose-derived mesenchymal stem cells with electromagnetic radiation, the frequency is 30 Hz, the intensity is 5 mt, for 7 consecutive days, once a day, 40 min each time, and at 37 ° C, 5 %CO 2 Cultivate in an incubator with high concentration and saturated humidity. Collect the cells at the end.

Embodiment 2

[0064] A method to rapidly increase the production of exosomes from adipose-derived mesenchymal stem cells:

[0065] 1. Prepare pre-activation solution: 0.2% laminin, 0.5% collagen, 0.2% fibronectin, 99.1% solvent, mix well and set aside.

[0066] 2. Take the fat after liposuction to separate and cultivate adipose-derived mesenchymal stem cells; take the P3-P5 generation of adipose-derived mesenchymal stem cells, count the cells and inoculate them in a culture flask at an equal density. The medium was Lonza serum-free medium + PALL's serum substitute + Gibco's glutamine mixture.

[0067] 3. Add 5 ng / mL pre-activation solution to the culture bottle, and treat the adipose-derived mesenchymal stem cells with electromagnetic radiation at a frequency of 70 Hz and an intensity of 1 mt for 7 consecutive days, once a day for 20 min each time, and incubate at 37°C, 5 %CO 2 Cultivate in an incubator with high concentration and saturated humidity. Collect the cells at the end.

Embodiment 3

[0069] A method to rapidly increase the production of exosomes from adipose-derived mesenchymal stem cells:

[0070] 1. Prepare pre-activation solution: 0.6% laminin, 0.3% collagen, 0.4% fibronectin, 98.7% solvent, mix well and set aside.

[0071] 2. Take the fat after liposuction to separate and cultivate adipose-derived mesenchymal stem cells; take the P3-P5 generation of adipose-derived mesenchymal stem cells, count the cells and inoculate them in a culture flask at an equal density. The medium was Lonza serum-free medium + PALL's serum substitute + Gibco's glutamine mixture.

[0072] 3. Add 3ng / mL pre-activation solution to the culture bottle, adipose-derived mesenchymal stem cells are treated with electromagnetic radiation, the frequency is 50Hz, the intensity is 3mt, for 7 consecutive days, once a day, each time for 30min, and at 37°C, 5 %CO 2 Cultivate in an incubator with high concentration and saturated humidity. Collect the cells at the end.

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Abstract

The invention discloses a method for quickly increasing the exosome yield of adipose tissue-derived stromal cells. The method is characterized in that the adipose tissue-derived stromal cells are subjected to electromagnetic radiation or/and laser radiation treatment for at least 7 days once every day and 20-40 minutes each time, and are cultured in an incubator at the temperature of 37 DEG C, theCO2 concentration of 5% and the saturated humidity; A pre-activation solution is adopted for protection treatment during electromagnetic radiation; and after the laser radiation treatment, a membraneglycoprotein repair solution is adopted for repair treatment. According to the method for rapidly increasing the exosome yield of the adipose tissue-derived stromal cells, the key factor for increasing the exosome yield is radiation and radiation condition control, exosomes can be effectively reserved in the cells through radiation, and are enriched in 7 days, and then the content of the exosomesis increased. Meanwhile, the pre-activation solution and the membrane glycoprotein repair solution are used for protecting the cells and preventing and repairing radiation damage; and the exosomes can be ensured to keep the original action effect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a method for rapidly increasing the exosome production of adipose-derived mesenchymal stem cells. Background technique [0002] Exosomes are lipid bilayer vesicles with a diameter of 40-100 nm. They were first proposed in 1983 by studying the vesicles secreted by sheep reticulocytes during maturation. They can carry proteins and lipids specifically expressed by cells. Substances such as cytoplasm and nucleic acid, and then directly transfer the carried substances to adjacent or distant cells through contact with cells. At present, a large number of studies have shown that exosomes derived from mesenchymal stem cells can play the role of tissue repair, immunosuppression, and immune regulation similar to mesenchymal stem cells through the intercellular transfer of protein and RNA components in their contents. And compared with cell therapy, exosomes are more stable ...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12N13/00
CPCC12N5/0667C12N13/00C12N2500/32C12N2500/34C12N2500/36C12N2500/90C12N2501/90C12N2501/998
Inventor 李栋董静戴晓宇刘抗赵莎莎张天友
Owner 重庆市铂而斐细胞生物技术有限公司
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