Umbilical cord mesenchymal stem cell exosome, preparation method and application of exosome to cosmetic
A technology of stem cells and exosomes, applied in the field of stem cells, can solve problems such as difficult to meet market demand
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Embodiment 1
[0029] Example 1: Isolation, cultivation, modification and transformation of umbilical cord mesenchymal stem cells and extraction of exosomes
[0030] 1. Experimental materials and instruments
[0031] The umbilical cord tissue was taken from healthy pregnant women who underwent cesarean section, stored at 4°C, and the umbilical cord mesenchymal stem cells were isolated and cultured within 24 hours. The puerpera herself signed the informed consent form, stating that the umbilical cords taken could only be used for scientific research.
[0032] The aseptic Petri dish with a diameter of 100 mm is a Corning product of the United States.
[0033] The cell incubator is a product of Thermo Fisher Scientific.
[0034] The flow cytometer is a product of BD Biosciences, USA.
[0035] Fetal bovine serum was the product of Gibco in the United States, α-MEM medium was the product of Hyclone, and trypsin was purchased from Beyond.
[0036] The fluorescently labeled flow antibodies CD14...
Embodiment 2
[0059] Example 2: Anti-aging activity test of exosomes
[0060] 1. Experimental materials and instruments
[0061] HFF-1 cells are products of Shanghai Huzhen Biotechnology Co., Ltd.
[0062] Fetal bovine serum was a product of Gibco, USA.
[0063] DMEM / F12 culture medium is a product of Sigma-Aldrich Company.
[0064] CCK-8 was purchased from Biyuntian; DMSO was a product of Sigma-Aldrich Company.
[0065] Cell culture box and microplate reader are products of Thermo Fisher Scientific.
[0066] 2. Experimental methods and results
[0067] 1. Cell culture
[0068] HFF-1 cells were cultured in DMEM / F12 medium with 10% fetal bovine serum at a temperature of 37°C and a concentration of 5% CO 2 Routine culture was carried out in a saturated humidity incubator, and when the cells grew to 85%, they were digested with trypsin and subcultured.
[0069] 2. Determination of cell proliferation activity by CCK-8
[0070] Take HFF-1 cells in the logarithmic growth phase, 1×10 per w...
Embodiment 3
[0075] Embodiment 3: A kind of anti-aging essence
[0076] Composed of the following raw materials: umbilical cord mesenchymal stem cell exosomes, 0.5%; sodium pyrrolidone carboxylate, 0.5%; glycerol, 3.5%; 1,3-propanediol, 3.5%; betaine, 1%; Glutathione, 0.05%; Disodium EDTA, 0.05%; Lauramidopropyl Hydroxysultaine, 0.5%; Allantoin, 0.1%; Ascorbic Acid, 0.1%; Sodium Hyaluronate, 0.1% ; Xanthan gum, 0.5%; essence, 0.05%; the balance is deionized water.
[0077] Exosomes released by mesenchymal stem cells have the characteristics of stem cells and have multiple biological functions such as promoting tissue repair, immunosuppression, and neuroprotection. Although mesenchymal stem cells are considered to be the most powerful exosome-producing cells, when used as cosmetic raw materials, it is still difficult to meet market demand. At this time, either by changing the in vitro culture parameters to improve the ability of mesenchymal stem cells to secrete exosomes, or by transformi...
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