Human-derived adipose tissue-derived stem cell factor, preparation method and application thereof
A technology of mesenchymal stem cells and cytokines, which is applied in the field of human adipose-derived mesenchymal stem cell factors and their preparation, can solve the problems that the effects need to be further improved, achieve good market prospects, accelerate the growth of skin collagen cells, and increase collagen secretion.
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Embodiment 1
[0029] Embodiment 1 Preparation of adipose-derived mesenchymal stem cell factor of the present invention
[0030] 1. Source of fat
[0031] The donor signed the informed consent form for fat donation, and the donor's coagulation function, hepatitis C virus antibody, HIV Ⅰ / Ⅰ antibody, hepatitis B virus antigen and Treponema pallidum antibody were checked before collection. Separation of fat to ensure that the collected fat is free of virus contamination.
[0032] 2. Preparation method
[0033] Fat-improved medium and hypoxic culture group of the present invention:
[0034] 1. Obtaining the supernatant of human adipose-derived mesenchymal stem cells (AADSC) hypoxic culture
[0035] (1) Cell culture:
[0036] Use a straw to suck out the liquid in the lower layer of the collection bottle, leaving the yellow fat tissue, and be careful not to suck the yellow fat particles;
[0037] Add 30ml of normal saline, shake the collection bottle, wash the adipose tissue, and then let it ...
Embodiment 2
[0076] Embodiment 2 The preparation of mesenchymal stem cell factor of the present invention
[0077] 1. Take the cytokine concentrate prepared in Example 1, and adjust the concentration of the cytokine concentrate through repeated ultrafiltration, so that the EGF content in the concentrate is 6ug / ml.
[0078] 2. Add a certain amount of lyoprotectant to the cytokine concentrate to make the EGF content 0.5ug / ml, and adjust the pH value to 6.0.
[0079] Among them, the lyoprotectant contains the following components per 100ml: tocopherol 0.5g, glutathione 0.4g, human albumin 0.5ml, dextran 3.5g, trehalose 1.5g, mannitol 4g, hydroxyproline Acid 0.05g, Arginine 0.02g, Histidine 0.2g, Methionine 0.2g, Tyrosine 0.2g, Glycine 2g, Sodium Citrate 0.26g. The freeze-drying conditions are as follows: the temperature is at -25-40° C., the pressure is maintained at 10-30 Pa, and the time is 18-28 hours to obtain the cytokine freeze-dried powder.
[0080] 3. Activity determination of cytok...
Embodiment 3
[0084] Embodiment 3 The preparation of cytokine cosmetic or medicine of the present invention
[0085] Take the freeze-dried cytokine powder prepared in Example 2, add biopolysaccharide glue, collagen, folic acid, and sterile ultrapure water to make a composition, which can be added to conventional skin care products or used alone as a skin care product .
[0086] Wherein 10ml of the aqueous solution contains 0.05g of cytokine freeze-dried powder, 0.1g of biological polysaccharide glue, 0.3ml of collagen sterile solution and 0.05g of folic acid.
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