Umbilical cord mesenchymal stem cell factor freeze-dried powder and preparation method thereof

A technology of mesenchymal stem cells and freeze-dried powder, which is applied in the field of umbilical cord mesenchymal stem cell factor freeze-dried powder and its preparation, can solve the problems of complex protein components, microbial infection, and many invalid proteins, so as to achieve faster proliferation and improve secretion capacity , the effect of low immune rejection

Active Publication Date: 2019-04-09
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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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  • Umbilical cord mesenchymal stem cell factor freeze-dried powder and preparation method thereof
  • Umbilical cord mesenchymal stem cell factor freeze-dried powder and preparation method thereof
  • Umbilical cord mesenchymal stem cell factor freeze-dried powder and preparation method thereof

Examples

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preparation example Construction

[0030] The invention provides a method for preparing umbilical cord mesenchymal stem cell factors, comprising the following steps: (1) taking Wharton's jelly from the umbilical cord, and culturing in vitro to obtain umbilical cord mesenchymal stem cells, which are recorded as P0 generation; (2) Passaging for 3 to 5 times, that is, after P3 to P5 generations, culture in a hypoxic environment for 4 to 8 hours, and mechanically stretch the cells; (3) restore normal oxygen concentration, and continue to culture in a carbon dioxide incubator; ( 4) Collecting the cell supernatant, performing ultrafiltration and dialysis, and collecting the retentate to obtain the umbilical cord mesenchymal stem cell factor.

[0031] Specifically, the method of taking Wharton's jelly from the umbilical cord and culturing umbilical cord mesenchymal stem cells in vitro is as follows: take the Wharton's jelly from the umbilical cord, place it in a petri dish until it adheres to the wall, add culture medi...

Embodiment 1

[0043] This embodiment provides a freeze-dried powder of umbilical cord 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: 15g skimmed milk powder, 10g trehalose, 8g carboxymethylcellulose, 3g hydroxyethyl starch, 0.05g glutamine, 3g poloxamer.

[0046] (2) Isolation of umbilical cord mesenchymal stem cells

[0047] 1) Take out the umbilical cord in the transport solution in the ultra-clean bench, put it in a petri dish, and wash it several times with cleaning solution to remove the blood on the surface of the umbilical cord;

[0048] 2) Cut the cleaned umbilical cord into 2-3cm long pieces and put them on the petri dish, and wash them several times with cleaning solution one by one to remove the blood remaining inside;

[0049] 3) Each segment was washed several times with 0.9% sterile saline without antibiotics, a...

Embodiment 2

[0069] This embodiment provides a kind of umbilical cord mesenchymal stem cell factor freeze-dried powder and its preparation method. Compared with Example 1, in step (4), the preparation of umbilical cord mesenchymal stem cell factor: 2) P3~P5 cells Passed to the mechanical stretching cell culture box, when the cell growth density reached 50% to 65%, the cells were subjected to 37°C, 5% CO 2 , 5%O 2 After acting for 6 hours, continue to culture in a carbon dioxide incubator; that is, do not perform the operation of stretching the elastic membrane by 20% according to the scale on the wall of the culture box.

[0070] All the other steps are the same as in Example 1.

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Abstract

The invention provides an umbilical cord mesenchymal stem cell factor freeze-dried powder and a preparation method thereof. Umbilical cord mesenchymal stem cells are cultured by mechanical stimulationand hypoxia stimulation to promote secretion of stem cell factors, and reasonable combination of components such as skimmed milk powder, trehalose, carboxymethyl cellulose, hydroxyethyl starch, glutamine and poloxamer is used as a freeze-dried powder protective agent. The prepared expected mesenchymal stem cell factor freeze-dried 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 freeze-dried powder of umbilical cord 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 repair...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/52C12N5/0775C12P21/00A61K9/19A61K47/46A61K47/26A61K47/38A61K47/36A61K47/18A61K47/10A61K35/28A61P17/02
CPCA61K9/19A61K35/28A61K47/10A61K47/183A61K47/26A61K47/36A61K47/38A61K47/46A61P17/02C07K14/52C12N5/0665C12P21/00
Inventor 黄燕飞车七石刘少辉
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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