Lyophilized powder of human mesenchymal stem cell culture supernatant and preparation method thereof

A technology of mesenchymal stem cells and supernatants, applied in the field of stem cells, can solve problems such as hindering popularization and application, and short storage time

Active Publication Date: 2013-08-21
冯文峰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the liquid culture supernatant has a short storage time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Isolation and culture of adipose-derived mesenchymal stem cells

[0023] Aseptically absorb the fat extraction solution, wash it several times with PBS to remove the drugs and blood cells used in the liposuction operation; digest with 0.1% type I collagenase at 37°C for 1 hour, centrifuge at 400g for 5 minutes, discard the upper layer of fat, mix well and use Filter through a 200-mesh sieve, centrifuge the obtained cell suspension at 1200 r / min for 8 min, wash the cell pellet with PBS, and adjust the cell concentration to 1×10 9 cells / L, suspended in low-sugar DMEM culture medium containing 10% autologous serum (10% autologous serum + low-sugar DMEM + penicillin 100 U / mL + streptomycin 100 U / mL), inoculated in a 10 cm culture dish, and placed at 37 ° C, Volume fraction 5%CO 2 Cultivate in an incubator; after 48 hours, suck out the suspended cell liquid, replace with a new medium, and change the medium once every 3 days, and subculture when the cells reach more than...

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Abstract

The invention relates to lyophilized powder of human mesenchymal stem cell culture supernatant and a preparation method of the lyophilized powder. The preparation method comprises the following steps of: culturing a sub-culturing human mesenchymal stem cell by using a mesenchymal stem cell culture medium containing autologous serums; collecting a cell culture supernatant with low sub-culturing times; and preparing the collected supernatant into the lyophilized powder. The lyophilized powder prepared by the preparation method of the lyophilized powder of the human mesenchymal stem cell culture supernatant effectively preserves various cell factor mixtures with biological activities in the human mesenchymal stem cell culture supernatant, is capable of effectively solving the problem of short storage life of the supernatant and lays a foundation for an industrial application of the human mesenchymal stem cell culture supernatant.

Description

technical field [0001] The invention relates to stem cells, in particular to a freeze-dried powder of human mesenchymal stem cell culture supernatant and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are important members of the stem cell family, derived from the mesoderm and ectoderm in the early stages of development. Mesenchymal stem cells were originally discovered in the bone marrow, and have attracted increasing attention because of their multi-lineage differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and self-replication. Under specific induction conditions in vivo or in vitro, mesenchymal stem cells can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelial and other tissue cells. It has multi-directional differentiation potential and can be used as an ideal seed cell for the repair of tissue and organ damage cau...

Claims

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Application Information

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IPC IPC(8): A61K35/12A61K9/19C12N5/0775A61P17/00A61K35/28
Inventor 冯文峰佘志勇方海庆陈建兴
Owner 冯文峰
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