Concentrated adipose-derived stem cell culture supernatant preparation and preparation method and application thereof
A technology of adipose stem cells and culture supernatant, which is applied in the field of concentrated adipose stem cell culture supernatant preparations, can solve the problems of unreasonable methods of repairing scald wounds, concurrent infection, limited use, etc., to achieve efficient repair, strong secretion ability, Apply handy effects
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Embodiment 1
[0032] This embodiment provides a concentrated adipose stem cell culture supernatant preparation, which is mainly obtained by concentrating the supernatant of a cell culture medium cultured from adipose tissue-derived mesenchymal stem cells. The specific preparation method of the concentrated adipose stem cell culture supernatant preparation of this embodiment is as follows: take 40 ml of adipose tissue (the adipose tissue is derived from animals, preferably from humans), and add collagenase type I at a concentration of 0.1% to 0.5% to digest for 30 to 30 ml. After washing with normal saline for 60 min, centrifuge at 2000 rpm / min for 5-10 min, take the precipitated cells, wash and resuspend with normal saline, filter with a filter membrane with a diameter of 70 μm, and obtain adipose-derived stem cells by adding GMP-standard hrEGF, VEGF and bFGF (concentrations of 0.08-0.15 μg / mL, 0.05-0.11 μg / mL and 0.01-0.10 μg / mL respectively) were cultured in serum-free medium for adipose-d...
Embodiment 2
[0034] This embodiment provides a concentrated adipose stem cell culture supernatant preparation, the difference from Example 1 is that in the final step of the concentrated adipose stem cell culture supernatant preparation in this embodiment, the step of adding cytokines hrEGF, VEGF and bFGF is removed .
[0035]The specific steps of this embodiment are as follows: take 40ml of adipose tissue, add 0.1%-0.5% type I collagenase to digest for 30-60min, wash with normal saline, centrifuge at 2000rpm / min for 5-10min, take Cells were precipitated, washed and resuspended with normal saline, and filtered with a filter membrane with a diameter of 70 μm to obtain adipose-derived stem cells. 0.01~0.10μg / mL) of adipose-derived stem cell serum-free medium for culturing; the adipose-derived stem cell serum-free medium in this embodiment was cultured in CO 2 The culture is carried out in an incubator with a temperature of 37° C. in a humid sterile gas environment with a content of 5%. The...
Embodiment 3
[0037] This embodiment provides a concentrated adipose stem cell culture supernatant preparation, the difference from Example 1 is that the GMP standard hrEGF, VEGF and bFGF added at the end of the finished product preparation are different from Example 1. Present embodiment 3 is divided into two examples, respectively as follows:
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