Preparation method of adipose-derived stem cell multi-cell active factor freeze-dried powder

An adipose stem cell and active factor technology, which is applied in the field of preparation of adipose stem cell multicellular active factor freeze-dried powder, can solve the problems of difficult to guarantee product uniformity, low content of cell active factor, low stem cell activity, etc., and achieve large-scale production. , reduce unfavorable factors, and promote the effect of cell proliferation

Pending Publication Date: 2020-09-04
成熙(上海)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, when conventional culture methods are used for culture, the activity of stem cells is low, the content of secreted cell activity factors is very small, the yield is low, the storage time is short, and it is necessary to collect multi-generation cell culture supernatants, the efficiency is low, and the uniformity of the product is low. Difficult to guarantee

Method used

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  • Preparation method of adipose-derived stem cell multi-cell active factor freeze-dried powder
  • Preparation method of adipose-derived stem cell multi-cell active factor freeze-dried powder
  • Preparation method of adipose-derived stem cell multi-cell active factor freeze-dried powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A method for preparing the freeze-dried powder of multi-cell active factors of fat stem cells, comprising the steps of:

[0035] S1: Collection of human adipose tissue:

[0036] A physical examination was performed on the donor, and then under local anesthesia, the fat was obtained by tumescent liposuction, and the fat and water were filtered out through a 500 μm filter hole to obtain light yellow fat.

[0037] S2: Primary isolation, culture and passage of adipose-derived stem cells:

[0038] (1) Wash the fat obtained in step S1 with PBS solution for several times until the cleaning solution is clear and translucent, and the fat is clean and light yellow, and then place the obtained fat in 0.2%-0.4% mixed collagenase ( In the mixture of type I collagenase and type VI collagenase), digest at 37°C with uniform shaking for 30-40 minutes, then add 0.25% Trypsin-EDTA digestion solution to it, shake evenly at 37°C After digestion for 10-15 minutes, add trypsin inhibitor and m...

Embodiment 2

[0052] Selection of Coating Reagents for Cell Culture Flasks

[0053] Coating the cell culture flask can enhance the cell adhesion rate and increase the proliferation efficiency. In this example, on the basis of Example 1, the culture flasks were not coated (Method 1), coated with gelatin (Method 2), coated with Laminin-111 (Method 3), and coated with Laminin-521 (Method 4), refer to Example 1 for other operating steps, respectively test the number of cells in the subculture process, and detect the cell activity factors in the finally prepared lyophilized powder. The experimental results are shown in Table 1 and Table 2.

[0054] Table 1 Effect of selection of cell culture flask coating reagent on cell proliferation

[0055]

[0056] The influence of the selection of table 2 cell culture bottle coating reagent on the content of cell viability factor

[0057]

[0058] The experimental results in Table 1 and Table 2 show that in Method 4, when the cell culture flask is c...

Embodiment 3

[0060] Effect of Oxygen Concentration on Secretion of Cell Activity Factors During Cell Anaerobic Culture

[0061] In this example, on the basis of Example 1, the oxygen concentration during the anaerobic culture of the cells during the large-scale expansion of the intermediate mesenchymal stem cells and the collection of the supernatant in step S3 is regulated respectively:

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Abstract

The invention discloses a preparation method of adipose-derived stem cell multi-cell active factor freeze-dried powder. The preparation method comprises the following steps: S1, collecting adipose tissue of a human body; S2, conducting primary separation, culture and passage of adipose-derived stem cells; S3, carrying out large-scale amplification on mesenchymal stem cells and collecting supernate; S4, preparing a concentrated stock solution of stem cell active factors; and S5, preparation of freeze-dried powder. The multi-cell active factor freeze-dried powder prepared by the preparation method disclosed by the invention includes a large number of cellular active factors, and is safe and reliable to use, free of any adverse reaction, high in yield, good in activity, easy to store and capable of remarkably promoting healing of deep cell tissues of damaged skin and fading scars and acne marks.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a preparation method of freeze-dried powder of adipose stem cell multicellular activity factor. Background technique [0002] Adipose-derived stem cells (ADSCs) are currently widely used in the field of regenerative medicine research. Compared with bone marrow stem cells and umbilical cord stem cells, adipose stem cells have the advantages of abundant raw materials, convenient material acquisition, and strong paracrine function, especially its strong paracrine function. There are about 800 kinds of cell growth factors in the human body, and ADSC can secrete 765 of them. It is superior to other mesenchymal stem cells and adult mesenchymal stem cells such as bone marrow and umbilical cord. [0003] After the discovery of the excellent paracrine function of adipose-derived stem cells, it was widely used in the medical cosmetology industry, and was gradually favored by other ind...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K9/19A61P17/02C12N5/0775
CPCA61K35/28A61K9/19A61P17/02C12N5/0667C12N2509/00C12N2533/52C12N2500/02
Inventor 唐健韩瑛璐罗平妃
Owner 成熙(上海)生物科技有限公司
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