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Stem cell preparation for skin beauty and preparation method thereof

A kind of stem cell preparation and stem cell technology, which is applied in the fields of skin care and beauty, can solve the problems of not being suitable for mass production, high requirements for culture conditions, and the inability to continue subculture of differentiated cells, so as to facilitate long-term storage, solve the problem of short storage time and multiple times of passage increased effect

Inactive Publication Date: 2017-02-22
JILIN TUO HUA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Related literature has reported that the use of hypoxic culture can increase the content of individual cytokines to a certain extent, but it requires a long-term hypoxic process, which requires high culture conditions, and with common culture methods, stem cells differentiate after multiple passages. Once differentiated, it no longer has the ability to secrete stem cell-related factors, and the differentiated cells cannot continue to be subcultured. The above problems are not suitable for mass production

Method used

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  • Stem cell preparation for skin beauty and preparation method thereof
  • Stem cell preparation for skin beauty and preparation method thereof
  • Stem cell preparation for skin beauty and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Preparation of stem cell preparation and freeze-dried product thereof by umbilical cord mesenchymal stem cells

[0047] 1. Acquisition and culture of primary umbilical cord mesenchymal stem cells

[0048] With the informed consent of the puerpera, the tests of hepatitis B virus antigen, hepatitis C surface antibody, HIV, Treponema pallidum antibody, etc. were negative, and a sterile neonatal umbilical cord was obtained.

[0049] Put the aseptically collected neonatal umbilical cord into physiological saline, soak it in 75% ethanol for 2 minutes, then wash it with PBS, remove the remaining blood in the blood vessel, and cut the umbilical cord into 1 mm pieces 3 Evenly spread the tissue pieces in culture dishes (Corning, Cat. 017) ɑ-MEM culture medium (Hyclone; product number SH30265.01)), at 37°C, 5% CO 2 (v / v) culture in an incubator for 3-5 days, and the grown cells are primary umbilical cord mesenchymal stem cells.

[0050] 2. Expansion and subculture of...

Embodiment 2

[0058] Example 2: Preparation of stem cell preparation and freeze-dried product thereof by bone marrow mesenchymal stem cells

[0059] 1. Acquisition and cultivation of primary bone marrow mesenchymal stem cells Collect 15ml of bone marrow from volunteers under aseptic conditions, anticoagulate with heparin, add 10ml of normal saline, mix well, slowly add into a 50ml centrifuge tube along the tube wall with a 5ml syringe, Add 15ml of Percoll cell separation medium (solarbio, product number P8370) into the centrifuge tube in advance, density gradient centrifugation, 2000r / min, centrifuge, 30min, collect the intermediate buffy coat cells in a sterile centrifuge tube, then add 30ml of normal saline to wash the cells, Centrifuge at 1500r / min for 10min, discard the supernatant, repeat washing twice, and finally add 20ml of ɑ-MEM culture solution containing 2% serum substitute, at 37°C, 5%CO 2 (v / v) culture in an incubator for 3-5 days, and the grown cells are primary umbilical cord...

Embodiment 3

[0061] Example 3: Preparation of stem cell preparation and freeze-dried product thereof by adipose-derived mesenchymal stem cells

[0062] After informed consent, the fat was obtained from female abdominal liposuction patients. 70ml of fat was washed twice with an appropriate amount of saline to remove some blood cells, and then an equal volume of 1.5% type I collagenase (solarbio, product number C8140) was added and placed in a constant temperature water bath at 37°C. Shake and digest for 40 minutes, remove the lower layer of mononuclear cells into a 50ml centrifuge tube, add 30ml of normal saline to wash the cells for 3 times, and finally add 20ml of ɑ-MEM culture medium containing 2% serum substitute, at 37°C, 5% CO 2 Culture in an incubator, and replace the medium or subculture depending on the growth status of the cells. Other preparation steps are the same as in Example 1.

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Abstract

The invention provides a stem cell preparation for skin beauty and a preparation method thereof, wherein the method comprises: a) providing stem cells acquired from a mammal animal; b) subjecting the stem cells to primary culture; c) subjecting the stem cells to subculture in a stem cell medium with vitamin A; d) changing with phenol-red-free culture liquid with astragaloside, and continuously culturing under hypoxic condition; e) subjecting the stem cells and the culture liquid to ultrasonic disruption and centrifuging, collecting supernate, and filtering to remove bacteria; f) detecting total protein concentration of the supernate, and adjusting the total protein concentration of the supernate. By using the preparation method of the stem cell preparation for skin beauty, the key problems of conventional culture methods, such as low in-vitro culture cell quantity and low factor content are solved.

Description

technical field [0001] The invention relates to the fields of beauty and skin care, in particular to a stem cell preparation for beauty and skin care and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are a kind of seed cells with self-renewal and multi-directional differentiation potential. Under different induction conditions, they can not only differentiate into bone, cartilage, fat, muscle and other mesoderm cells, but also It can differentiate across the germ layer to form neurons, glial cells, skin cells of the ectoderm, and vascular endothelial cells of the endoderm and other tissue cells. In addition to having multi-directional differentiation potential, MSC can also secrete a variety of cytokines, such as EGF, KGF, VEGF, angiopoietin-1 (angiopoietin-1, Ang-1), bFGF, IGF- 1. Cell active substances such as Wnt1 / 3A / 5A, HGF, PDGF-BB and EPO are well known to promote angiogenesis, tissue cell biosynthesis and energy metabolism,...

Claims

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Application Information

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IPC IPC(8): A61K8/64A61K38/17A61P17/02A61P17/16A61P3/10C12N5/0775
CPCA61K8/64A61K38/1709A61K2800/10C12N5/0663C12N5/0665C12N5/0667C12N5/0668C12N2500/34C12N2500/38
Inventor 田娜张石林尹晓光
Owner JILIN TUO HUA BIOTECH
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