Proliferation accelerant and application thereof

A technology of accelerator and freeze-dried excipient, which is applied in the field of biology to achieve the effects of increasing content, improving production efficiency, and promoting expression and secretion

Active Publication Date: 2018-07-10
SHENZHEN HUADA GENE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are currently no relevant literature reports on the research of bitter melon saponins in inducing skin fibroblasts to synthesize and secrete collagen and cytokines and other biologically active substances.
At the same time, loofah extract can improve the activity of human skin fibroblasts and has obvious antioxidant effect, but there is no report on the application of luffa saponin to the cultivation and induction of skin fibroblasts

Method used

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  • Proliferation accelerant and application thereof
  • Proliferation accelerant and application thereof
  • Proliferation accelerant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1: Primary isolation, culture, subculture and identification of skin fibroblasts

[0091] (1) Primary isolation, culture and passage of skin fibroblasts

[0092] After wiping the skin with iodine and subcutaneously injecting a small amount of anesthetic, the skin tissue is taken by professional medical personnel with a medical skin sampler to obtain a skin block with a size of about 3mm in diameter. Put the skin in a PBS solution containing 1000U / mL penicillin and 1000μg / mL streptomycin, fully wash off the blood on the tissue block, cut off the peduncle tissue appropriately and cut the remaining skin tissue into 4-8 pieces, and then Transfer the tissue pieces to a culture dish containing PBS solution containing 100 U / mL penicillin and 100 μg / mL streptomycin and wash thoroughly; cut up the skin tissue pieces and transfer them to a 15mL centrifuge tube, add 1-2mL with a mass volume ratio of 0.25 % neutral protease at 37°C for 1-3 hours, separate the epidermis and...

Embodiment 2

[0095] Example 2: Preparation process of fibroblast extract freeze-dried powder

[0096] (1) Preparation of Human Skin Fibroblast Extract Solution

[0097] Human skin fibroblasts in good growth state were subcultured and expanded in T-175 culture flasks, and the culture medium was phenol red-free DMEM high-glucose medium containing 10% (v / v) serum substitute (penicillin 100 U / mL, strepto Mycin 100 μg / mL, pH7.2). After the cell confluence reaches 80%-90%, use trypsin digestion solution (0.25% (w / w) trypsin + 0.02% (w / w) EDTA) to digest the cells, press 5000-6000cell / cm 2 Subculture. When the cells were subcultured to the P6-P8 generation, the subculture was stopped, and the skin fibroblast supernatants of each passage were collected and stored at -80°C until processing. Trypsinization was used to collect the skin fibroblasts by centrifugation for ultrasonication. The ultrasonication conditions were: 4°C, 400W, ultrasonication for 3s, interval of 4s, 99 cycles. After the son...

Embodiment 3

[0100] Example 3: Effects of luffa saponin and bitter melon saponin composition on skin fibroblast collagen synthesis and cytokine secretion

[0101] After identification, the P3-P5 skin fibroblasts in good growth state were taken for culture, and the cells were collected to adjust the cell density to 5×10 4 cell / mL, inoculate cells into a six-well plate, 2 mL per well. After the cells were cultured for 24 hours, the original culture medium was removed, and the skin fibroblast culture medium containing the combined stimulator of loofah saponin and bitter melon saponin was added as shown in Table 1 for culture. After the cells were cultured for 3 days, the cell culture supernatants of each group were collected. Using human type I collagen (Collagen I), human transforming growth factor (TGF-β), human platelet-derived growth factor (PDGF-AB), human basic fibroblast growth factor (bFGF) ELISA kits, The contents of collagen and major cytokines secreted by human skin fibroblasts w...

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Abstract

The invention relates to a proliferation accelerant and application thereof, in particular to a fibroblast proliferation accelerant capable of promoting fibroblast proliferation and fibroblast extractand fibroblast extract freeze-dried powder prepared through fibroblasts. The proliferation accelerant comprises loofah saponin and/or momordica saponin. A loofah saponin and momordica saponin combined inducer is adopted for culture of skin fibroblasts, so that synthesis and secretion of bioactive substances derived from the skin fibroblasts are promoted effectively, content of active ingredientsin cell derivatives is increased, and production efficiency and biologic effect of the skin fibroblast derived cell derivatives.

Description

technical field [0001] The invention relates to the field of biology, relates to a proliferation promoter and its application, in particular to a fibroblast proliferation promoter capable of promoting fibroblast proliferation, and a fibroblast extract and a fibroblast extract prepared from fibroblasts Extract lyophilized powder. Background technique [0002] At present, all kinds of skin care products are mainly made of chemical ingredients, supplemented by plant extracts and other nutrients. Although they have certain effects, they have great limitations due to safety or effectiveness issues. Therefore, "green skin care products" based on safety, functionality and nature have become the current development trend. [0003] Using skin fibroblasts as the basic resource to exert the biologically active substances contained in the cells (such as collagen, cytokines, amino acids, vitamins, etc.), has been widely studied and applied by professionals in the fields of medicine or b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K35/33A61K9/19A61K8/98A61P17/02A61Q19/00A61Q19/02
CPCA61K8/98A61K9/19A61K35/33A61K47/42A61Q19/00A61Q19/02C12N5/0656C12N2500/34
Inventor 岳建辉何娜刘亚琼康晖张曦
Owner SHENZHEN HUADA GENE INST
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