Preparation method of striae gravidarum-removing cytokine freeze-dried powder in mesenchymal stem cell culture supernate
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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.
Inactive Publication Date: 2019-08-02
江苏全能干细胞生物工程有限公司
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Commonly used techniques include laser or photon irradiation, microcrystalline grinding, fruit acid peeling, abdominoplasty, etc., but due to insufficient effects or obvious side effects, it is not accepted by most people
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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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Abstract
The invention discloses a preparation method of striae gravidarum-removing cytokine freeze-dried powder in mesenchymal stem cell culture supernate. The preparation method of the striae gravidarum-removing cytokine freeze-dried powder in the mesenchymal stem cell culture supernate comprises the following steps of preparing primary adipose mesenchymal stem cells and primary umbilical cord mesenchymal stem cells, and building a cell bank; thawing a P1 generation of adipose mesenchymal stem cells and a P1 generation of umbilical cord mesenchymal stem cells, adding normal saline, conducting centrifugation, collecting cell sedimentation, inoculating a serum-free medium against the cell sedimentation, and conducting plate culture; 4-5 days after plate culture is conducted, when the cell density reaches 70%, collecting culture supernate, adding EDTA recombinant trypsin digestive cells, culturing the mesenchymal stem cells to a P5 generation, and collecting the culture supernate; adding normalsaline into a cell factory of the P5 generation of cells, after repetitive freeze-thawing, mixing the cells and the P2-P5 mesenchymal stem cell culture supernate, conducting centrifugation, and removing sedimentation; and adding trehalose, mannitol and hyaluronic acid, and conducting ultra-low-temperature vacuumizing freeze-drying to obtain the cytokine freeze-dried powder.
Description
technical field [0001] The invention relates to the technical field of stem cells, in particular to a preparation method of cytokine freeze-dried powder in the culture supernatant of mesenchymal stem cells for eliminating stretch marks. Background technique [0002] The formation of stretch marks is mainly due to the influence of hormones during pregnancy. The swelling of the abdomen causes the elastic fibers and collagen fibers of the skin to be damaged or broken to varying degrees due to external force. The skin becomes thinner and thinner. Pink or fuchsia wavy pattern of varying lengths. After childbirth, these patterns will gradually disappear, leaving white or silvery white shiny scar lines, that is, stretch marks. Medical beauty technology can reduce or eliminate stretch marks by increasing the production of collagen, improving the color and thickness of the skin epidermis, and removing the skin at the stretch marks. Commonly used techniques include laser or photon i...
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