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Mesenchymal stem cell composite active factor freeze-dried powder as well as preparation method and application thereof

An active factor, stem cell technology, used in cell culture active agents, biochemical equipment and methods, animal cells, etc., can solve the problems of component extraction, purification can not be completely removed, high risk, allergies and other problems, to promote division and Proliferate, promote metabolism, restore skin elasticity

Active Publication Date: 2020-04-10
广州赛加生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most media contain commercially available fetal bovine serum or chemical components that are not suitable for the human body. For example, fetal bovine serum may carry unknown pathogens or foreign proteins such as prion protein. Once it enters the human body, it may cause infection and allergy and other adverse events, the risk is high, and these components cannot be completely removed in the process of extraction, purification, and concentration, so it is very restrictive in clinical application

Method used

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  • Mesenchymal stem cell composite active factor freeze-dried powder as well as preparation method and application thereof
  • Mesenchymal stem cell composite active factor freeze-dried powder as well as preparation method and application thereof
  • Mesenchymal stem cell composite active factor freeze-dried powder as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0052] In Example 2, the commercially available serum-free complete medium for adipose stem cells was purchased from Celera Stem Cell Technology Co., Ltd., and the product number is S-10-012;

[0053] MEM-α medium was purchased from Shanghai Yuanpei Biotechnology Co., Ltd., the product number is L562KJ.

Embodiment 1

[0054] Example 1: Extraction and phenotypic identification of adipose stem cells

[0055] Take excess fat from healthy volunteers after liposuction, add excess sterile double-antibody-containing PBS buffer, soak for 10-15 minutes, then centrifuge at 2000 rpm for 10 minutes to remove blood and PBS buffer, repeat once. Transfer the obtained fresh adipose tissue to a 50ml centrifuge tube, add an equal volume of 0.075% collagenase and 0.25% trypsin (1:1), and digest in a water bath at 37°C for 40-50min. After the liquid becomes chylus, centrifuge at 1000rpm for 5min, and filter with a 200-mesh cell sieve, transfer the filtrate to a culture bottle, add the first medium, and transfer to 5% CO 2 , 95% air, incubate in a cell incubator at 37°C, observe the growth status of the cells the next day, and perform half or full replacement of the medium. When the cell fusion reaches 80%, use 0.25% trypsin ~ EDTA to digest the cells , to obtain primary adipose stem cells.

[0056] Wherein, ...

Embodiment 3

[0063] Example 3: Effect of pre-coating treatment on the attachment performance of human-derived adipose-derived stem cells

[0064] Take 10ml of 10μg / ml fibronectin and 0.001g / ml polyvinyl alcohol (PVA) mixed equal volume pre-coating solution, gently add to the 75cm along the wall of the culture flask 2 In the culture bottle, gently tap the culture bottle with your fingers to remove air bubbles to ensure that the pre-coating solution fully covers the bottom wall of the culture bottle, transfer it to a 4°C refrigerator, and place it for 30 minutes for later use.

[0065] Take 10ml of 10μg / ml fibronectin pre-coating solution, and gently add it along the wall of the culture flask to 75cm 2 In the culture bottle, gently tap the culture bottle with your fingers to remove air bubbles to ensure that the pre-coating solution fully covers the bottom wall of the culture bottle, transfer it to a 4°C refrigerator, and place it for 30 minutes for later use.

[0066] Take 10ml of 0.001g / m...

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Abstract

The invention relates to mesenchymal stem cell composite active factor freeze-dried powder as well as a preparation method and application thereof. In the preparation process of the composite active factor freeze-dried powder, a specific pre-coating solution is used for treating a cell culture container; and mesenchymal stem cells are cultured by adopting a first culture medium and a second culture medium under specific culture conditions, so the mesenchymal stem cells can be still kept at a normal state in the processes of hunger culture and induction and secrete more active factors. The composite active factor freeze-dried powder prepared by the invention can promote cell division and cell proliferation, repair damaged skin or skin with thin congenital cuticle, tighten the skin and reduce wrinkles; moreover, the immunoregulation function of the skin is activated, the metabolism of the skin is promoted, and an anti-aging effect is achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to freeze-dried powder of mesenchymal stem cell compound active factor and its preparation method and application. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of mesoderm and ectoderm derived from prenatal embryonic development stages, and exist in various tissues and organs, such as bone marrow, adipose tissue, muscle tissue, lung, liver, pancreas, etc. and synovium, etc. MSCs have multi-directional differentiation ability and paracrine function. Under the action of different inducing factors, they can differentiate into adipocytes, chondrocytes, and city cells; they can also secrete a variety of cytokines, including chemokines, adhesion molecules, etc. In recent years, studies have shown that various biologically active factors secreted by stem cells can be used as a new type of cell-free biological regulation mode to promote or complete intercellular signa...

Claims

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Application Information

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IPC IPC(8): A61K38/40A61K9/19A61P39/06A61K8/02A61K8/99A61K8/98A61K8/64A61K8/67A61Q19/08C12N5/0775A61K38/06A61K31/375A61K35/28
CPCA61K8/022A61K8/64A61K8/676A61K8/981A61K8/982A61K8/99A61K9/0014A61K9/19A61K31/375A61K35/28A61K38/06A61K38/40A61K2800/805A61Q19/08A61P39/06C12N5/0667C12N2500/24C12N2500/38C12N2501/105C12N2501/11C12N2501/115C12N2501/15C12N2501/91C12N2501/998A61K2300/00
Inventor 曾伙妹洪军辉马钦海陈永康谢文凤
Owner 广州赛加生物科技有限公司
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