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Cell growth factor freeze-dried powder, preparation method and application

A growth factor, freeze-dried powder technology, applied in the field of stem cells, can solve the problems of allergy or malabsorption, inability to exert the effect of cytokines, rejection, etc. good protective effect

Inactive Publication Date: 2018-06-15
深圳市新一仑生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing technology on the market is mainly to extract cytokines and prepare them into liposome wraps, or injections, or smears, because the biological activity of cytokines will gradually disappear over time due to improper storage. characteristics, so it cannot exert the real effect of cytokines
Based on the above reasons, directly using MSC as the source of cytokines to prepare freeze-dried powder is regarded as the most simple and effective way to protect the activity of cytokines. For example, patent CN106367386 A adopts the method of directly freeze-drying it for preservation, and CN 106109496 A is based on this Add trehalose as a protective agent to further protect the activity and properties of cytokines, but trehalose or other exogenous additives may cause rejection, resulting in allergies or malabsorption

Method used

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  • Cell growth factor freeze-dried powder, preparation method and application
  • Cell growth factor freeze-dried powder, preparation method and application
  • Cell growth factor freeze-dried powder, preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of cytokine freeze-dried powder

[0034] A: Isolation and culture of mesenchymal stem cells

[0035] 1) Take back the fresh umbilical cord with a sterile bottle (200ml normal saline + a gentamicin);

[0036] 2) Separate the Wharton's jelly around the blood vessel, and cut it into pieces;

[0037] 3) Add 2mg / ml type Ⅰ collagenase (about 15ml);

[0038] 4) Add 200 μl of double antibody and 200 μl of amphotericin B, mix well and place in an incubator (37°C) overnight;

[0039] 5) After taking it out, add 10ml (depending on the digestion of collagenase) trypsin, and put it in the incubator (37°C) for digestion for 30min;

[0040] 6) Take it out, dilute it 4-6 times with normal saline, and then centrifuge at 3000rpm for 15-20min;

[0041] 7) Discard the supernatant, add medium to suspend, transfer to 2-3 petri dishes (about 20ml / dish), add 1ml additive, 200μl double antibody and 200μl amphotericin B to each dish, and place in the incubator to cultivate;

[...

Embodiment 2

[0101] Comparison of cytokine activity under different storage conditions

[0102] Under different storage conditions of the freeze-dried powder, the content of cytokines epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), and exosome (exosome) was determined. The experimental results are shown in the attached Figure II shown.

[0103] The experimental results show that the storage temperature of the cell growth factor freeze-dried powder is below 4°C and -20°C to ensure its good activity.

[0104] Activity comparison of different cytokine products stored at 4°C

[0105] The lyophilized powder of the present invention, the lyophilized powder of Comparative Example 1, the lyophilized powder of Comparative Example 2, and the finished stem cell mask on the market were stored at 4°C for different periods of time to determine the content of cytokines EGF and bFGF:

[0106] 1) Place the four kinds of stem cell test products in...

Embodiment 3

[0114] skin test experiment

[0115] Skin test experiment: The stem cell factor freeze-dried powder of the present invention and the finished stem cell facial mask on the market can improve human facial skin;

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Abstract

The invention provides cell growth factor freeze-dried powder, a preparation method and application. The cell growth factor freeze-dried powder is prepared by freezing cell culture supernate obtainedfrom subculture of umbilical cord mesenchymal stem cells, and is used as a protection agent after human albumin is added. The preparation method comprises the following steps: A, separating mbilical cord mesenchymal stem cells; B, performing subculture on the mbilical cord mesenchymal stem cells in a culture medium; C, collecting mbilical cord mesenchymal stem cell culture supernate of the step B,adding mannitol and human albumin, sucking the cell culture supernate into a centrifugal tube, placing in a low-temperature refrigerator or a liquid nitrogen tank, performing cryopreservation till asolid, completely freezing and solidifying the culture medium, putting the solid frozen culture medium into a cryopreservation bottle, performing freezing drying in a machine for 12-24 hours so as toobtain freeze-dried powder, and performing cold storage on the freeze-dried powder. As the human albumin is adopted as a protection agent, irritation and other rejection reactions are not easily caused, the powder can be relatively easily absorbed, a skin oxidation resistance effect is achieved, and skin aging can be further retarded.

Description

Technical field: [0001] The invention relates to the technical field of stem cells, in particular to a cell growth factor freeze-dried powder, a preparation method and an application. Background technique: [0002] MSC is currently the most researched stem cell. Many clinical trials have confirmed that MSC can treat a variety of diseases and has a good effect. Cell growth factor is a multifunctional and powerful cytokine that promotes the metabolism of fibroblasts and the production of collagen. Formation plays an important function. The content in the body is very small, but it has high biological activity; it can promote and regulate the division, proliferation and differentiation of skin cells, and play a biological role in regulating the physiological functions and metabolic activities of various cells. The repair and growth of cells makes the skin smooth and plump, which can be called our youth factor. The existing technology on the market is mainly to extract cytokin...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K9/19A61K47/42A61P17/18A61P17/00C12N5/0775
Inventor 杨前刁波张宜袁紫林陈力王刚王丽萍
Owner 深圳市新一仑生物科技有限公司
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