Chitosan microsphere composition loaded with stem cell active factors and preparation method and application thereof
A technology of chitosan microspheres and active factors, applied in skin care preparations, pharmaceutical formulations, cosmetic preparations, etc., can solve the problems of low transdermal absorption and delivery efficiency of stem cell active factor skin care products, and achieve continuous protection effect , Enhanced utilization, skin roughness improvement effect
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[0032] First aspect, a kind of preparation method of the chitosan microsphere composition that is loaded with stem cell activity factor provided by the present invention, comprises the steps:
[0033] (1) Stem cells are cultured and amplified with clinical-grade serum-free medium, and the culture supernatant is collected after regular medium change and passage, and the stem cell active factor solution is obtained after removing cells and cell debris;
[0034](2) Add lactic acid to the stem cell activating factor solution obtained in step (1), then add chitosan oligosaccharides, and form a polycation mixed solution after stirring, wherein the volume ratio of the lactic acid to the stem cell activating factor solution is 0.5 ~5:100, the ratio of the mass of chitosan to the volume of the stem cell activating factor solution is 0.5~1:100 (g / mL);
[0035] (3) Add the polyanion solution into the polycation mixed solution obtained in step (2) under stirring, and react to obtain the c...
Embodiment 1
[0057] Example 1 Preparation of Umbilical Cord Mesenchymal Stem Cell Active Factor Solution
[0058] (1) Isolation of umbilical cord mesenchymal stem cells: place the umbilical cord that has passed the quality inspection (sterile, negative for infectious disease, and relatively hypertrophic in appearance) in a petri dish, and peel off the outer skin and blood vessels under aseptic conditions; Wash off the residual blood with the gel-like substance, and cut the tissue into a clean culture dish; wash with DMEM medium, add mesenchymal stem cell serum-free medium to resuspend, add it to a T75 culture bottle and culture it in a cell culture incubator ;
[0059] (2) Expansion of mesenchymal stem cells: the primary cells are subcultured after they grow to 50-80% confluent, and then subcultured every 2-3 days until P5;
[0060] (3) Collect the supernatant of stem cell activity factor: P1-P5 generation cells collect the cell culture supernatant before each passage, mix and filter (0.4...
Embodiment 2
[0072] The present embodiment provides a kind of preparation method of the chitosan microsphere suspension that is loaded with stem cell active factor, comprises the steps:
[0073] According to the method of Example 1, the stem cell activating factor solution was obtained, and 1 L of the resulting stem cell activating factor solution was measured. Under aseptic conditions, 10 ml of lactic acid was added to the stem cell activating factor solution and magnetically stirred at a speed of 100 rpm for 2 minutes, followed by adding 5g of chitosan oligosaccharides was stirred magnetically at a speed of 100rpm for 10min to dissolve chitosan oligosaccharides to obtain a mixed solution; under magnetic stirring conditions of a speed of 100rpm, 100ml of sodium tripolyphosphate Solution (mass concentration is 1wt%) is uniformly sprayed in the mixed solution of gained, after preparation, continue to stir 30min and make microsphere harden, the solution that prepares is the chitosan microsphe...
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