Repair mask containinghigh concentration human mesenchymal stem cell secretion active factors and preparation method thereof
A technology of mesenchymal stem cells and active factors, which is applied in the field of repairing masks containing high concentrations of human mesenchymal stem cells secreting active factors and its preparation, can solve the problems of repairing facial skin that have not been reported, so as to enhance skin repair ability and promote blood vessel repair. Effects of regeneration and wound healing
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[0022] The preparation method of the repairing mask containing high-concentration human mesenchymal stem cell-secreted active factor of the present invention comprises the following steps:
[0023] (1) Human mesenchymal stem cell culture: P2-P6 human mesenchymal stem cells were selected from the cell bank for resuscitation, the inoculation batches were recorded as the first batch, the second batch, and the third batch, and the inoculation density was the maximum fusion 50%-60%, 35%-45%, 20%-30% of the temperature; the cells are inoculated and cultured in a carbon dioxide incubator at a temperature of 37°C and a volume ratio of carbon dioxide of 5%;
[0024] (2) When the fusion degree of the first batch of cells reaches 70%-80%, wash the cells twice with normal saline, and perform starvation culture with compound electrolyte injection;
[0025] (3) After 12-24 hours of starvation culture, the first batch of cell confluence reaches 90%-95%, collect the cell culture supernatant, ...
Embodiment 1
[0039] (1) Culture of human umbilical cord mesenchymal stem cells: P4 human umbilical cord mesenchymal stem cells were selected from the cell bank for recovery, the total number of cells was 4.2*10 7 . The inoculation batches were recorded as the first batch, the second batch, and the third batch, and the inoculation density was 60% of the maximum fusion degree (the number of inoculated cells per bottle: 9*10 6 , inoculate three bottles of cells), 40% (the number of cells inoculated in each bottle: 6*10 6 , two bottles of inoculated cells), 20% (number of inoculated cells: 3*10 6 , inoculate a bottle of cells), the culture system is 40ml. The complete medium used for culturing the cells is phenol red-free DMEM / DF12 and 10% fetal calf serum by volume, and the bottle for inoculating the cells is a T-175 culture flask. After the cell inoculation is completed, place it in a carbon dioxide incubator for cultivation at a temperature of 37°C and a carbon dioxide concentration of 5...
Embodiment 2
[0050] (1) Culture of human adipose-derived mesenchymal stem cells: P5 human adipose-derived mesenchymal stem cells were recovered from the cell bank, and the total number of cells was 3.84*10 7 . The inoculation batches were recorded as the first batch, the second batch, and the third batch, and the inoculation density was 50% of the maximum fusion degree (the number of inoculated cells per bottle: 8*10 6 , inoculate three bottles of cells), 35% (number of cells inoculated in each bottle: 5.6*10 6 , two bottles of inoculated cells), 20% (number of inoculated cells: 3.2*10 6 , inoculate a bottle of cells), the culture system is 40ml. The complete medium used for culturing the cells is AMEM without phenol red and 10% fetal bovine serum by volume, and the bottle for inoculating cells is T-175 culture flask. After the cell inoculation is completed, place it in a carbon dioxide incubator for cultivation at a temperature of 37°C and a carbon dioxide concentration of 5%;
[0051...
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