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A human-derived mesenchymal stem cell factor and its preparation method and use

A technology for stem cells and cytokines, which is applied in the field of human mesenchymal stem cell factors and their preparation, can solve the problems of low content of cytokines, low practical application value, loss of cytokines, etc., and achieves promotion of nutritional metabolism and good market prospects. , the effect of increasing collagen secretion

Active Publication Date: 2019-12-17
SICHUAN NEO LIFE STEM CELL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cytokine cosmetics on the market are generally manufactured by directly mixing stem cell culture supernatant or lyophilized into cosmetic matrix. The content of cytokines is low and contains a lot of sugar and other impurities. After using these cosmetics, it will Cause discomfort such as dry skin
[0004] There are also some reports on the extraction and purification of stem cell culture supernatant, but the effect is still not good. For example, the patent with the notification number CN102600057B discloses a preparation method of human placental stem cell extract freeze-dried powder. A single 3000D filter membrane is used for ultrafiltration interception, which removes some impurities, but the impurities such as sugar are still not removed, and it also causes a large loss of cytokines, poor repair effect, and also causes discomfort such as dry skin , the practical application value is low

Method used

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  • A human-derived mesenchymal stem cell factor and its preparation method and use
  • A human-derived mesenchymal stem cell factor and its preparation method and use
  • A human-derived mesenchymal stem cell factor and its preparation method and use

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1 The preparation of mesenchymal stem cell factor of the present invention

[0034] 1. The source of the umbilical cord

[0035] The parents of the fetus signed the informed consent form for umbilical cord donation, and the maternal antibodies to hepatitis C virus, HIV Ⅰ / Ⅱ, hepatitis B virus antigen and Treponema pallidum were checked prenatally. The umbilical cord and umbilical cord blood with qualified inspection results were collected at the same time of delivery. Umbilical cord blood is used to detect viruses to ensure that the collected umbilical cords are free of virus contamination.

[0036] 2. Preparation method

[0037] 1. Acquisition of human umbilical cord mesenchymal stem cell (UC-MSC) culture supernatant

[0038] (1) Under sterile conditions, cut 10 donated umbilical cords that passed the virus test into 1mm pieces with ophthalmic scissors 3 Tissue pieces of about size are then mixed well.

[0039] (2) Evenly spread the uniformly mixed tissu...

Embodiment 2

[0063] Embodiment 2 The preparation of mesenchymal stem cell factor of the present invention

[0064] 1. Take the cytokine concentrate prepared in Example 1, and adjust the concentration of the cytokine concentrate through repeated ultrafiltration, so that the EGF content in the concentrate is 0.4ug / ml.

[0065] 2. Add a certain amount of lyoprotectant to the cytokine concentrate to make the EGF content 0.3ug / ml, and adjust the pH value to 6.0.

[0066] Among them, the lyoprotectant contains the following components per 100ml: ascorbic acid (VC) 0.5g, human albumin 0.5ml, dextran 3.5g, trehalose 1.5g, glycine 0.04g, arginine 0.17g, glycine 2g, 0.26g of sodium citrate, and 15ml of tert-butanol.

[0067] The freeze-drying conditions are as follows: the temperature is at -25-40° C., the pressure is maintained at 10-30 Pa, and the time is 16-24 hours to obtain the cytokine freeze-dried powder.

[0068] 3. Activity determination of cytokine freeze-dried powder

[0069] As determ...

Embodiment 3

[0072] Embodiment 3 The preparation of cytokine cosmetic or medicine of the present invention

[0073] Take the freeze-dried cytokine powder prepared in Example 2, add hyaluronic acid, collagen, and sterile ultrapure water to make a composition, which can be added to conventional skin care products, or used alone as a skin care product.

[0074] 100ml of the aqueous solution contains 1.0g of cytokine freeze-dried powder, 0.5g of low molecular weight (3000D-10KD) hyaluronic acid, 2ml of sterile collagen solution, and 5g of water-soluble azone.

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Abstract

The invention discloses a method for separating and purifying a human-derived mesenchymal stem cell factor, which comprises the following steps: a. acquisition of cell culture supernate: taking human mesenchymal stem cells, culturing in a mesenchymal stem cell serum-free culture medium, and when the cell grows to the confluence of 75-85%, collecting the supernate; and b. filtration and concentration: filtering the supernate in the step a through a 0.22 mu m filter membrane, passing the supernate through a 50KD ultrafiltration membrane, collecting the dialysate, passing the dialysate through a 1KD ultrafiltration membrane, and collecting the trapped fluid to obtain the cell factor concentrated solution. The invention also discloses the mesenchymal stem cell factor prepared by the method and application thereof. The invention also discloses a cosmetic containing the mesenchymal stem cell factor. The method can effectively remove impurities in the mesenchymal stem cell culture solution; and the obtained cell factor has the advantages of high purity, high yield and favorable application prospects.

Description

technical field [0001] The invention belongs to the technical field of skin restoration, and in particular relates to a human-derived mesenchymal stem cell factor and its preparation method and application. Background technique [0002] In recent years, with the deepening of research on mesenchymal stem cells, its secreted factors have become a hot spot for researchers. Mesenchymal stem cells can secrete a variety of biologically active cytokines, which can effectively regulate body cell signal transduction, activate human stem cells, and then physiologically repair or replace damaged, diseased and aging cells in the body. Adding cytokines to beauty cosmetics not only has the moisturizing and whitening effect of general cosmetics, but also can repair damaged skin, eliminate skin wrinkles, shrink pores, improve complexion, etc., and has attracted more and more attention. [0003] Cosmetics containing cytokines are now on the market in many countries, especially in South Kore...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C07K14/475C07K1/34A61K8/98A61K8/64A61K35/28A61K38/18A61Q19/00A61Q19/08A61P17/02A61P17/00
CPCA61K8/64A61K8/982A61K35/28A61K38/18A61K47/42A61Q19/00A61Q19/08C07K14/475
Inventor 任莹莹金百华何进琼王荣邓云涛
Owner SICHUAN NEO LIFE STEM CELL BIOTECH
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