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Preparation method of freeze-dried powder containing stem-cell active factors

An active factor and freeze-dried powder technology, applied in the field of stem cell active factor preparation, can solve the problems that the content of factors in the supernatant of stem cells cannot be significantly increased, the content of secreted factors is low, and the moisture content of freeze-dried powder is high, so as to promote The effect of healing deep cells and tissues of damaged skin, lightening scars and acne marks, and promoting the new generation of subcutaneous blood vessels

Active Publication Date: 2018-11-06
廊坊康宝汇泰生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the medium is used for conventional culture, the activity of stem cells is low, the content of secreted factors is very small, the yield is low, and the storage time is short. Using some conventional methods, such as mechanical friction, ultraviolet radiation and starvation culture, can not significantly improve Factor content in stem cell supernatant
At present, there are also some patent disclosures on the preparation methods of stem cell active factors and freeze-dried powders. For example, Chinese patent application 201710015650.7 discloses a human adipose-derived mesenchymal stem cell factor and its preparation method and use. The concentrated The content of mesenchymal stem cell factors in liquid adipose tissue has been improved to a certain extent, and the quality is stable. The product prepared from the composition with other materials in a specific ratio has a certain ability to repair skin damage; Chinese patent application 201610680823.2 discloses a human The preparation method of the freeze-dried powder of the culture supernatant of mesenchymal stem cells and the prepared freeze-dried powder, which preserve the various biologically active cytokine mixtures in the culture supernatant of human mesenchymal stem cells to the greatest extent, are used in the The application of cosmetics and other aspects has laid the foundation; but when the stem cell active factor or its freeze-dried powder prepared by the method disclosed in the above patent is applied to cell repair, not only the repair effect is not good, but the wound healing rate after burns is low. , during the preparation process, there is also a large loss of cytokines, and the moisture content in the freeze-dried powder is high, which leads to the problem that the freeze-dried powder is not easy to store

Method used

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  • Preparation method of freeze-dried powder containing stem-cell active factors
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  • Preparation method of freeze-dried powder containing stem-cell active factors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 A kind of preparation method of stem cell active factor freeze-dried powder, comprises the following steps:

[0038](1) Detection of exogenous factors: ELISA diagnostic kit was used to detect Treponema pallidum, HIV, hepatitis B virus, hepatitis C virus, human cytomegalovirus, human Epstein-Barr virus and T-lymphoblastic leukemia virus on the aspirated adipose tissue supernatant. Wait for exogenous factor detection, and then judge the test results, so as to ensure that the above exogenous factors are not contained in the adipose tissue.

[0039] (2) Primary isolation, culture and subculture of adipose stem cells: First, take about 20ml of adipose tissue and put it in a 50ml centrifuge tube, dilute it, blow it away, and then centrifuge, carefully extract the sediment at the bottom, wash the adipose tissue, and then use 0.25% trypsin Digest the extracted bottom sediment with 1% collagenase digestion solution, then centrifuge again to absorb the cells and put ...

Embodiment 2

[0051] The only difference from Example 1 is that in the step (2) of the preparation method of the stem cell active factor freeze-dried powder, the digestion solution is used to digest the extracted bottom sediment, and the digestion solution is: 0.1 % trypsin and 0.5% collagenase digestion solution.

Embodiment 3

[0053] The only difference from Example 1 is that in the step (2) of the preparation method of the stem cell active factor freeze-dried powder, the digestion solution is used to digest the extracted bottom sediment, and the digestion solution is: 0.3 % trypsin and 2% collagenase digestion solution.

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Abstract

The invention relates to the field of technical research of stem cells and provides a preparation method of freeze-dried powder containing stem-cell active factors excreted by human mesenchymal stem cells. Provided human adipose-derived stem cell raw materials are sufficient in source, safe and convenient to extract and safe and reliable in use and do not produce any untoward effect. The preparation method specifically comprises the following steps of exogenous factor detection, primary isolation, culture and cell passage of adipose-derived stem cells, identification of mesenchymal stem cells,collection of supernate, preparation of the freeze-dried powder, detection on the supernate and active factors of the stem cells of the freeze-dried powder and solvent preparation and filling. The prepared freeze-dried powder is easy to store and transport compared with the supernate, the activity of the factors is effectively saved, the freeze-dried powder can be applied to the beauty industry,the cell factor content is high, the moisture content in the freeze-dried powder is low, and accordingly the freeze-dried powder is easy to store. The freeze-dried powder prepared by adopting the preparation method is adopted after scalding, and the heal rate is high when the compound prepared from biological polysaccharide gel, collagen, folic acid and sterile ultrapure water is applied to wounds.

Description

technical field [0001] The invention belongs to the field of biotechnology medicine, relates to the field of stem cell technology research, in particular to a preparation method and application of a stem cell active factor secreted by human adipose mesenchymal stem cells. 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 mesench...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C07K14/52G01N33/68A61K35/28A61P17/02
CPCA61K35/28A61P17/02C07K14/52C12N5/0667G01N33/6863
Inventor 李玉才吴炳元成芳苏晓华
Owner 廊坊康宝汇泰生物技术有限公司
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