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Method for preparing adipose-derived stem cells with function of high collagen expression and product prepared from adipose-derived stem cells

An adipose stem cell and collagen technology, which is applied to the preparation of adipose stem cells with high collagen expression and the field of its products, can solve the problems of unbearable pain, slow growth, and inability to meet the needs of multiple beauty treatments.

Pending Publication Date: 2018-11-06
岳萍
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when adipose stem cells are cultured and proliferated in vitro, the growth rate is slow, reaching 10 8 The above number of cells needs more than 20 days, and it cannot meet the needs of multiple beauty treatments
Many beauticians may need liposuction for the second beauty treatment, but this kind of pain is very painful, and it is difficult for ordinary people to accept
Even with local anesthesia, the pain is unbearable afterwards
[0004] To sum up, the currently commonly used adipose stem cell culture method obtains a limited number of adipose stem cells, and the proliferation and growth of adipose stem cells is slow. 8
Moreover, the subculture and expansion of adipose stem cells should not exceed 5 generations, otherwise the adipose stem cells are prone to differentiation and apoptosis, and the functions of cell proliferation and repair are also significantly reduced, and the cells with stem cell characteristics are significantly reduced, and the activity of adipose stem cells after cosmetic treatment cannot be guaranteed.

Method used

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  • Method for preparing adipose-derived stem cells with function of high collagen expression and product prepared from adipose-derived stem cells
  • Method for preparing adipose-derived stem cells with function of high collagen expression and product prepared from adipose-derived stem cells
  • Method for preparing adipose-derived stem cells with function of high collagen expression and product prepared from adipose-derived stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Preparation of human adipose stem cells

[0056] Cosmetology volunteers obtained 30 mL of adipose tissue or isolated adipose tissue from the abdomen or thigh by liposuction (and signed an informed consent form with them), and washed them with 50 mL of normal saline repeatedly for 5 times to remove as much oil and blood cells as possible in the fat. Using 50 mL of 0.25% (mass volume ratio) trypsin (Gibco) and 0.1% (mass volume ratio) type I collagenase (Sigma, USA) at a ratio of 1:1, digest in a 37°C incubator for 60 minutes, every 15 Vibrate with a vortex shaker for 2 minutes; after digestion, centrifuge at 1500rpm for 10 minutes, absorb the upper layer of fat, add physiological saline to resuspend, and filter with a 100μm cell sieve (BD Company, USA); centrifuge the cell suspension at 1000rpm for 10 minutes, and use physiological Add OriCell after resuspending in saline and washing twice TM Serum-free medium (Lonza Company of the United States) (containing 1...

Embodiment 2

[0059] Example 2 Preparation of Recombinant Human Type IV Collagen α3 Coated Plate and Cell Culture Flask

[0060] Weigh 1 mg of recombinant human type IV collagen α3, fully dissolve it with 200 mL 1×PBS (pH=7.4), and filter and sterilize with a 0.45 μm filter to prepare 5 μg ml of recombinant human type IV collagen α3 coating solution. Add 500μL to each well of a 6-well plate, 90mm add 2mL, 75cm 2 Add 4mL and 175cm 2 Add 4 mL of recombinant human type IV collagen α3 coating solution to the culture bottle, incubate overnight at 4°C or 1 hour at 37°C, and then coat in a cell culture vessel. After coating, suck off the collagen solution, ventilate and dry in a biological safety cabinet or ultra-clean bench, seal and store in a refrigerator at 4°C after drying.

Embodiment 3

[0061] Example 3 Analysis of Proliferation Activity of Human Adipose Stem Cells

[0062] The adipose stem cells prepared in Example 1 and the adipose stem cells prepared in the comparative example were taken for proliferative activity analysis. The comparative examples were cultured to obtain adipose stem cells according to the method in the article published in the Journal of Low Temperature Biology by Park S et al. in Feb. Ahn CW, Kim H. Fetal bovine serum-free cryopreservation methods for clinical banking of human adipose-derived stem cells. Cryobiology. 2018 Feb 13.pii: S0011-2240(17)30288-2.)

[0063] Cell proliferation activity was detected on day 1, day 2, day 3, day 4, and day 5, respectively, by a Bio-Rad cell counter in the United States according to the instruction manual. from figure 1 It can be known that the adipose stem cells prepared in the present invention have a faster cell proliferation rate than those prepared in the comparative example, and there is a s...

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Abstract

The invention provides a method for preparing adipose-derived stem cells with the function of high collagen expression. The method comprises the following steps: a recombinant human type-IV collagen alpha 3 coated cell cultivator is used, under joint culture of recombinant human epithelial growth factors, recombinant human basic fibroblast growth factors, vitamin E and argania spinosa kernel oil,the adipose-derived stem cells with the function of high collagen expression are prepared and grow from adipose, 1*10<9> or more adipose-derived stem cells can be obtained by culturing 30 mL of adipose-derived cells within 14 days, mass cell quantity required by clinical aesthetic surgery can be met, secondary liposuction can be avoided, and pain caused by secondary liposuction can be relieved. According to the method, the invention further provides a product prepared from the adipose-derived stem cells with the function of high collagen expression. The product can be applied to preparation and growth of the adipose-derived stem cells with the function of high collagen expression.

Description

technical field [0001] The invention relates to a preparation method of adipose stem cells highly expressing collagen and a product prepared by the adipose stem cells highly expressing collagen obtained by the method. In particular, the present invention relates to cell culture plates or dishes coated with recombinant human type IV collagen α3 and cell culture flasks coated with recombinant human type IV collagen α3. Preparation and growth of adipose stem cells with high collagen protein expression from fat under the joint co-cultivation of fiber growth factor, vitamin E and argan tree kernel oil. The fat stem cells obtained in the invention can be used in medical cosmetology, including the fields of skin wrinkle removal, freckle removal, breast enhancement and the like. Background technique [0002] With the improvement of people's quality of life, there is a new pursuit of health and beauty. Medical cosmetology has achieved unprecedented development at present, and the m...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2500/38C12N2500/30C12N2501/115C12N2501/11C12N2533/54
Inventor 不公告发明人
Owner 岳萍
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