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Method for preparing human adipose-derived stem cell culture solution and application thereof in cosmetic products

A technology of human adipose stem cells and adipose stem cells, which is applied in the field of stem cell technology, can solve the problems that adipose stem cells do not have the same standard and affect the quality and quantity of adipose stem cells, and achieve the effects of shortening digestion time, promoting paracrine function, and avoiding damage

Inactive Publication Date: 2019-01-11
深圳市乔水生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, there is no uniform standard for in vitro culture methods of adipose stem cells, which affects the quality and quantity of adipose stem cells finally obtained

Method used

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  • Method for preparing human adipose-derived stem cell culture solution and application thereof in cosmetic products
  • Method for preparing human adipose-derived stem cell culture solution and application thereof in cosmetic products
  • Method for preparing human adipose-derived stem cell culture solution and application thereof in cosmetic products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1.1 Preparation of adipose stem cell culture medium (ADSC-CM)

[0033] This example will describe the process of preparing ADSC-CM in detail in the following content.

[0034] Step a: Obtain normal adipose tissue subcutaneously, about 500 mg, rinse with calcium and magnesium-free Hank's solution, use ophthalmic scissors to cut as much as possible the small blood vessels visible to the naked eye in the adipose tissue, wash the blood, cut the adipose tissue into pieces, transfer it into a large glass tube, add 0.25vol% trypsin (sigma) and 0.1vol% collagenase (type I, sigma) (volume ratio 1:1), 37 ℃ constant temperature shaking table shaking digestion for 20 minutes; at this time the liquid surface is divided into 3 layers: the upper layer is yellow Oily fat cell layer, the middle layer is adipose tissue layer, and the lower layer is liquid containing mononuclear cells;

[0035] Step b: Aspirate the lower layer of liquid and transfer it to a centrifuge tube containing com...

Embodiment 2

[0050] This example is set to be compared with Example 1 to demonstrate the promoting effect of the technical solution of Example 2 on the paracrine function of ADSCs.

[0051] 2.1 The effect of partial pressure of oxygen on the paracrine function of ADSCs

[0052] In this part, the partial pressure of oxygen during the culture of ADSCs was changed, except that other processing steps were kept the same as in Example 1. The culture atmosphere conditions of each group are as follows:

[0053] 1%O 2 Pretreatment group: volume fraction 1% O 2 +5%CO 2 +94%N 2 ;

[0054] 3%O 2 Pretreatment group: volume fraction 3% O 2 +5%CO 2 +92%N 2 ;

[0055] 5%O 2 Pretreatment group: volume fraction 5% O 2 +5%CO 2 +90%N 2 ;

[0056] 10%O 2 Pretreatment group: volume fraction 10% O 2 +5%CO 2 +85%N 2 ;

[0057] Control group (embodiment 1): volume fraction 5% CO 2 +95% air, ie normal / routine cell culture conditions.

[0058] ELISA was used to measure the amount of paracrine f...

Embodiment 3

[0080] Studies have shown that insulin-like growth factor, epidermal growth factor, transforming growth factor β, interleukin 1 and tumor necrosis factor α secreted by ADSCs can improve skin fibroblast damage caused by UVB, strengthen fibroblast proliferation and The ability to synthesize collagen, among which transforming growth factor beta stimulated fibroblasts most strongly in vitro.

[0081] 3.1 Preparation of whitening and anti-wrinkle beauty emulsion

[0082] The human-derived adipose-derived mesenchymal stem cell culture fluid obtained in Example 1 is used to prepare whitening and wrinkle-removing cosmetic emulsion, and the specific steps are as follows:

[0083] (1) Preparation of Matrix Group Emulsion

[0084] The mass percent between each component is as follows:

[0085]

[0086] Preparation of water phase: Accurately weigh deionized water, heat to about 60°C, add methylparaben, wait until completely dissolved, add disodium edetate, sodium hyaluronate, mix wel...

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Abstract

The invention discloses a method for preparing human adipose-derived stem cell culture solution and an application thereof, comprising the following steps: digesting human adipose tissue by using trypsin and collagenase, dividing the solution into three layers, taking the lower layer, and preparing adipose stem cell precipitate; terminating L-The digestion after being added in DMEM complete mediumto carry out primary culture; culturing primary adipose-derived stem cells (ADSCs) in hypoxic atmosphere with O2 partial pressure of 1 to 5% and CO2 partial pressure of 5% and ultraviolet irradiationof 10 to 30 mJ / cm2; culturing the cells in 1*10-7mol / L insulin to obtain adipose-derived stem cell culture; centrifuging an adipose-derived stem cell culture solution, filtering, and removing the adipose-derived stem cells to obtain a filtrate which is the adipose-derived stem cell culture solution. The culture method can promote the paracrine of adipose-derived stem cells, and the obtained adipose-derived stem cell culture medium contains a plurality of paracrine components, and the content is very high. As the active substance, the adipose stem cell culture solution can be adde to skin careand beauty products, so that the skin care and beauty product has the effect of whitening, removing scar, anti-wrinkle, and the like.

Description

technical field [0001] The invention relates to the application of stem cell technology, in particular to a method for preparing human adipose stem cell culture fluid and its application in cosmetic products. Background technique [0002] Adipose-derived stem cells (ADSCs) are mesenchymal stem cells obtained from adipose tissue. Since adipose-derived stem cells (ADSCs) were first isolated and named from adipose tissue in 2001, ADSCs have a wide range of sources, convenient materials, and tissue The advantages of small damage and rapid expansion have become ideal tools in cell therapy and tissue engineering applications. Many studies have found that ADSCs can play a significant role in the repair of damage in vivo and anti-aging; part of the realization of its biological role is through the multi-directional differentiation ability, ADSCs can differentiate into fat, bone, cartilage, muscle, tendon, etc. Through the paracrine function, ADSCs can synthesize and secrete a varie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K8/99A61Q19/10A61Q19/02
CPCA61K8/99A61Q19/02A61Q19/10C12N5/0667C12N2500/02C12N2501/33
Inventor 魏色东
Owner 深圳市乔水生物科技有限公司
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