Preparation method of striae gravidarum-removing cytokine freeze-dried powder in mesenchymal stem cell culture supernate
A technology of stem cells and culture supernatant, applied in the field of stem cells, can solve the problems of insufficient effect and obvious side effects, and achieve the effect of optimizing the types of cytokines, improving the storage time and ensuring the activity.
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Embodiment 1
[0037] Example 1: A method for preparing the freeze-dried powder of cytokines from the culture supernatant of mesenchymal stem cells for eliminating stretch marks, according to the following steps:
[0038] a) The umbilical cord tissue was obtained from the obstetrics and gynecology department of a tertiary hospital, washed with normal saline until there were no obvious red blood cells, the umbilical cord was cut into small pieces of about 2 cm with scissors, and the tissue was cut along the venous vessel, the venous vessel wall and arterial vessel were removed with tissue forceps, and strips were taken Wharton's jelly, cut into small pieces of 1-2mm3 with scissors, inoculated in serum-free culture medium, and half changed the medium every 5 days, on the 15th day of culture, the cell density reached 50%, added EDTA recombinant trypsin to digest Cells, after washing with normal saline, centrifuge to collect the cell pellet, add freezing solution to freeze the cells at -80°C;
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Embodiment 2
[0061] Example 2: A method for preparing the freeze-dried powder of cytokines from the culture supernatant of mesenchymal stem cells for eliminating stretch marks, according to the following steps:
[0062] The steps from a) to j) were the same as those in Example 1. After collecting the supernatant of the cultured P5 generation cells, 1 L of normal saline was added to the cell factory, and the cells were repeatedly frozen and thawed at -20° C. to lyse the cells.
[0063] f) Centrifuge the cell lysate at 5000rpm for 30min to remove the cell debris, mix evenly with the collected cell culture supernatant, filter the mixed culture supernatant with a 0.22um capsule filter, and add seaweed in proportions of 5%, 8%, and 1% respectively Sugar, mannitol, and hyaluronic acid are mixed evenly to prepare a freeze-dried mixture.
[0064] g) Divide the freeze-dried mixed stock solution into vials, pre-freeze quickly, and set the pre-freeze temperature to -40°C.
[0065] h) When the pre-fr...
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