Small pet eye drops containing human mesenchymal stem cell excreta and preparation method thereof
A mesenchymal stem cell and secretion technology, which is applied in the field of eye drops containing human mesenchymal stem cell secretion and its preparation, can solve the problems of harsh survival conditions, discounts, and easy-to-death living cells of active cells, so as to shorten the healing time , Eliminate inflammation, promote the effect of repair
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[0020] The preparation method of the small pet eye drops containing human mesenchymal stem cell secretion of the present invention comprises the following steps:
[0021] (1) Mesenchymal stem cell culture: P2-P6 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%;
[0022] (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;
[0023] (3) After 12-24 hours of starvation culture, the first batch of cell confluence reaches 90%-95%, collect the cell culture supernatant, centrifuge at 3000-4500g for 5-8min to ...
Embodiment 1
[0032] (1) Human umbilical cord mesenchymal stem cell culture: P2 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 , three bottles of cells inoculated), 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 volume ratio concentration of carbon...
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