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Mesenchymal stem cell extract and its use

A technology of mesenchymal stem cells and extracts, applied in the field of mesenchymal stem cell extracts and their uses, can solve the problems of affecting the appearance of the skin, trauma, and limited application

Active Publication Date: 2015-08-19
GWOXI STEM CELL APPL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the current application of cell therapy still relies on injection into the body
The most commonly used method is to inject a certain amount of stem cells under the skin. This invasive treatment method not only has certain risks, but also will cause many traumatic holes, which will affect the appearance of the skin
In addition, this traditional cell therapy uses living cells, and its safety must be strictly evaluated before injection, and when administered to humans, the source of the cells must be human stem cells, so its applicability is quite limited
[0007] In view of the above-mentioned extracellular matrix supplementation therapy, the limited effect and side effects of botulinum toxin, and the potential risks and limitations of cell therapy, it is urgent to develop a simple method that can effectively repair skin aging

Method used

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  • Mesenchymal stem cell extract and its use
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  • Mesenchymal stem cell extract and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Isolation and cultivation of mesenchymal stem cells

[0046] Experiment (1): Isolation of mesenchymal stem cells from tissue

[0047]Cut adipose tissue from different human bodies (such as NR0, NR1, NR2, NR3, NR4, AD03, AD61, BN08, and MS14) into small pieces (about 0.5 cm x 0.5 cm) with surgical scissors, and the cut adipose tissue Place in a 50ml centrifuge tube, wash with 25ml of 1X phosphate buffered saline (PBS) solution containing 2% penicillin-streptomycin, and centrifuge (room temperature, 400g, 5 minutes). After centrifugation, the supernatant was removed, washed with 25 ml of 1XPBS solution containing 2% penicillin-streptomycin, and centrifuged (room temperature, 300g, 5 minutes). After centrifugation, the supernatant was removed, and 20 ml of 1X PBS solution containing 0.4 mg / ml collagenase type IV (purchased from Invitrogen) was added, and placed on a shaker for 1 hour (37°C, 40 rpm). After the type IV collagenase has fully interacted with the a...

Embodiment 2

[0051] Example 2: Analysis of Chemokines in Mesenchymal Stem Cells

[0052] (1) Preparation of whole-cell RNA from mesenchymal stem cells

[0053] For the mesenchymal stem cells provided in Example 1, wash once with 0.1 millimolar PBS solution, add 350 microliters of RLT buffer (QIAGEN, CA, USA) containing 1% β-mercaptoethanol, and mix Uniformly, then add an equal volume (ie, 350 microliters) of 70% ethanol, mix evenly, transfer the mixture to RNeasy mini colum (QIAGEN, CA, USA), and perform centrifugation (12000rpm, 15 seconds). After centrifugation, remove the supernatant, then add 700 μl of RW1 buffer (QIAGEN, CA, USA), and perform centrifugation (12000 rpm, 15 seconds); after centrifugation, remove the supernatant, and then add 500 μl of RPE buffer (QIAGEN, CA, USA), perform centrifugation (12000rpm, 15 seconds); remove the supernatant after centrifugation, repeat the step of RPE buffer once, and perform centrifugation (12000rpm, 1 minute) to remove residual reagents. Mo...

Embodiment 3

[0061] Example 3: Preparation of mesenchymal stem cell extract

[0062] First, suspend the mesenchymal stem cells provided in Example 1 with trypsin-diamineethylenetetraacetic acid (Trypsin-EDTA), then transfer them to a centrifuge tube, and perform centrifugation (room temperature, 400g, 5 minutes), After centrifugation, remove the supernatant, and then add an appropriate amount of pre-prepared protein lysis buffer [including NP40 lysis buffer (iNtron biotechnology, Korea), phenylmethylsulfonyl fluoride (PMSF), protease inhibitors, β-glycerol Phosphate, and sodium vanadium, the dosage ratio is 96:1:1:1:1], and repeated suction with a micropipette to break the cells, placed at 4 ° C for 20 minutes, and then centrifuged (4 °C, 12000 to 13000 rpm, 10 minutes), after centrifugation, the supernatant (ie, mesenchymal stem cell extract) was collected for use in the following examples.

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Abstract

A mesenchymal stem cell extract and its use are provided, wherein the mesenchymal stem cell extract comprises a trophic factor(s), such as bone morphogenetic protein-7 (BMP-7), stromal cell-derived factor-1 (SDF-1), vascular endothelial growth factor (VEGF), C-X-C chemokine receptor type-4 (CXCR4), brain-derived neurotrophic factor (BDNF), and / or interleukin-17 (IL-17), and wherein the extract is especially suitable for repairing skin aging.

Description

technical field [0001] The present invention relates to an extract extracted from mesenchymal stem cells and the application of the extract, especially to the application of the extract in making cosmetics or medicines for repairing skin aging, especially to using the extract to promote the production of hyaluronic acid in the skin , promote elastin production in the skin, promote collagen production in the skin, promote reticulin production in the skin, stimulate cell proliferation at the application site, stimulate cell secretion of chemokines (trophic factors) at the application site, and / or stimulate endogenous Hematopoietic stem cells gather to the site of administration and proliferate. When the mesenchymal stem cell extract of the present invention is used in combination with a heating operation, it can especially provide an excellent effect of repairing skin aging. Background technique [0002] The skin is the largest organ of the human body. It covers the surface o...

Claims

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Application Information

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IPC IPC(8): A61K8/64A61K38/21A61K38/20A61K38/19A61K38/18A61K38/17A61Q19/08A61Q7/00A61P17/00A61P17/14
CPCA61F7/08A61F2007/0062A61F2007/0003A61N5/0625A61K8/981A61Q19/08A61K2800/70A61K8/64A61F7/02A61F2007/0088A61K2800/805A61K2800/86A61K2800/88A61P17/00A61P17/14A61Q7/00
Inventor 林欣荣韩鸿志邱紫文林珀丞
Owner GWOXI STEM CELL APPL TECH CO LTD
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