Adipose-derived mesenchymal stem cell factor lyophilized powder and preparation method thereof

A technology of mesenchymal stem cells and freeze-dried powder, which is applied in the field of adipose-derived mesenchymal stem cell factor freeze-dried powder and its preparation, can solve the problems of low efficiency of stem cell factors, complex protein components, and many ineffective proteins, so as to improve secretion capacity and source Widespread, active-boosting effects

Active Publication Date: 2019-04-16
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The efficiency of secreting stem cell factors by cultivating stem cells in ordinary methods is low, and the bioactive peptide solution of stem cells in a liquid state obtained by secreting is stored for a short time at room temperature, which is not conducive to transportation, and the concentration is also low
In addition, stem cell bioactive peptides and stem cell seeds from allogeneic sources have been screened, but there is still a risk of microbial infection
There are also some products that are directly obtained by directly lysing stem cells or other active cells. The protein composition is complex. Although the concentration is high, there are many ineffective proteins, which will also cause metabolic stress on the human body.

Method used

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  • Adipose-derived mesenchymal stem cell factor lyophilized powder and preparation method thereof
  • Adipose-derived mesenchymal stem cell factor lyophilized powder and preparation method thereof
  • Adipose-derived mesenchymal stem cell factor lyophilized powder and preparation method thereof

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preparation example Construction

[0029] The invention provides a method for preparing adipose-derived mesenchymal stem cell factor, which comprises the following steps: (1) taking adipose tissue, enzymatically digesting and culturing it to obtain adipose-derived mesenchymal stem cells; After passages P3 to P5, culture in a hypoxic environment for 1 to 3 hours, and mechanically stretch the cells; (3) restore the normal oxygen concentration and continue the culture; (4) collect the cell supernatant, ultrafiltration dialysis, and collect The retentate is the adipose-derived mesenchymal stem cell factor.

[0030] Wherein, the adipose tissue is the human adipose tissue obtained by the cosmetology department of the hospital with the consent of the patient.

[0031] Specifically, the isolation and purification of adipose-derived mesenchymal stem cells includes 1) aseptically extracting adipose tissue from itself, soaking it in sterile saline containing double antibodies, and sending it to the laboratory; , washed 3...

Embodiment 1

[0043] This embodiment provides a kind of freeze-dried powder of adipose-derived mesenchymal stem cell factor and its preparation method, the specific steps are as follows:

[0044] (1) Preparation of lyoprotectant

[0045] Each 100mL cytokine concentrate contains the following components: 5g polyethylene glycol, 3g konjac sugar, 0.5g proline, 1g glutathione, 0.025g sodium benzoate, 0.25g sodium dodecylsulfonate, 0.5g ectoine.

[0046] (2) Isolation and purification of adipose-derived mesenchymal stem cells

[0047] 1) Aseptically extract your own adipose tissue, soak it in sterile saline containing double antibodies, and send it to the laboratory;

[0048] 2) Take out the adipose tissue, wash it with sterile normal saline for 3 times, transfer the adipose tissue to a cell culture dish, add an appropriate amount of 0.25% EDTA-Tripsin+0.5% protease compound enzyme, and put it in a 37°C cell incubator for digestion for 3 hours;

[0049] 3) Stop the digestion of the serum-c...

Embodiment 2

[0061] This embodiment provides a kind of freeze-dried powder of adipose-derived mesenchymal stem cell factor and its preparation method. Compared with Example 1, (3) Preparation of adipose-derived mesenchymal stem cell factor: 2) transfer P3-P5 cells to mechanical In the pulling cell culture box, when the cell growth density reaches 50% to 65%, the cells are subjected to 37°C, 5% CO 2 , 5%O 2 After acting for 2 hours, continue to culture in a carbon dioxide incubator; that is, do not perform the operation of stretching the elastic membrane by 15% according to the scale on the wall of the culture box.

[0062] All the other steps are the same as in Example 1.

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Abstract

The invention provides adipose-derived mesenchymal stem cell factor lyophilized powder and a preparation method thereof. According to the method, an adipose-derived mesenchymal stem cell is cultured by adopting conditions of mechanical stimulation and hypoxic stimulation, the secretion of stem cell factors is promoted, the reasonable cooperation of polyethylene glycol, konjak sugar, proline, glutathione, sodium benzoate, sodium dodecyl sulfate and tetrahydro-2-pyrimidinone is adopted to serve as a lyophilized powder protective agent, and the prepared adipose-derived mesenchymal stem cell factor lyophilized powder has good stability, stem cell factor content and activity.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to a kind of freeze-dried powder of adipose mesenchymal stem cell factor and a preparation method thereof. Background technique [0002] During the stable growth process of stem cells, many kinds of cytokines are secreted into the medium supernatant in the form of protein peptides. By collecting and treating the supernatant after culturing stem cells, a large number of cytokines can be obtained through a series of treatments. These people Derived factors include EGF (epidermal growth factor), VEGF (vascular endothelial growth factor), HGF (liver growth factor), PDGF (platelet-derived factor), bFGF (fibroblast growth factor) and the like. Most of these factors have biological activities related to skin growth, blood vessel growth, and subcutaneous tissue regeneration. They can effectively regulate body cell signal transduction, activate human cells, and then physiologically repai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/475C07K1/34A61K38/18A61K9/19A61K47/10A61K47/36A61K47/22A61K47/18A61K47/12A61K47/20A61P17/02
CPCA61K9/19A61K38/18A61K47/10A61K47/12A61K47/183A61K47/20A61K47/22A61K47/36A61P17/02C07K14/475
Inventor 黄燕飞车七石刘少辉
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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