Application of umbilical cord mesenchymal stem cell conditioned medium
A technology of conditioned medium and stem cells, applied in the field of conditioned medium of umbilical cord mesenchymal stem cells, can solve the problems of inconvenient storage and transportation, immune rejection, etc., and achieve low impact of environmental factors, sufficient tissue sources, product stability and safety. high sex effect
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Embodiment 1
[0047] The preparation of embodiment 1.UCMSC-CM
[0048] UCMSC (third generation) was provided and identified by Beijing Stem Cell Bank, Institute of Zoology, Chinese Academy of Sciences. Seed UCMSCs in a 100mm culture dish, add 10ml UCMSC medium (DMEM / F12, 10% FBS, GlutaMAX, penicillin-streptomycin, 4ng / ml basic FGF) for culture, when the cell confluency reaches 80%, discard the culture culture medium, washed 3 times with sterile PBS, added 6ml of serum-free medium (DMEM / F12) to each 100mm culture dish, placed the cells in a 37°C, 5% CO2 cell incubator for 24 hours, and collected the culture medium after the culture was over. And centrifuge, 1500rpm, centrifuge for 5 minutes to remove cell debris, aliquot into 1ml centrifuge tubes, store at -80°C for later use, and the collected conditioned medium UCMSC-CM comes from the 4th-6th generation UCMSC.
Embodiment 2
[0049] Example 2. Establishment of Ovarian Injury Model in Mice
[0050] The mouse ovary at 5 days after birth (P5) is mainly composed of primordial follicles and primary follicles, which are extremely sensitive to toxic agents, so mice at 5 days after birth are ideal for studying the mechanism of chemotherapy drug-induced death of immature follicles time. P5 mice were given a single intraperitoneal injection of cisplatin, the dose of cisplatin injection was 5 mg / kg body weight, and the mice in the control group were injected with the same amount of normal saline. The mice were killed by cervical dislocation 4 days after injection of cisplatin or normal saline, and the ovaries were collected. And perform fixed embedding section ( figure 1 A).
[0051] Results: Compared with the control group, the primordial follicles in the ovaries of the cisplatin treatment group were significantly reduced ( figure 1 B). Therefore, in the following in vivo experiments in mice, we selected...
Embodiment 3
[0052] Example 3. The protective effect of UCMSC-CM on ovarian damage caused by cisplatin in mice
[0053] In order to explore the protective effect of human umbilical cord mesenchymal stem cell conditioned medium on cisplatin-induced ovarian damage in mice, CD-1 mice born 5 days old (P5) (purchased from Speifu (Beijing) Laboratory Animal Technology Co., Ltd. , mother mice feeding) 80, weighing about 2.8g body weight. Divide P5 mice into four groups, blank control group: continuous intraperitoneal injection of common medium for 4 days; cisplatin group: a single intraperitoneal injection of 5 mg / kg body weight of cisplatin on the first day of treatment, and then continuous intraperitoneal injection of common medium 3 days; conditioned medium group: continuous intraperitoneal injection of conditioned medium for 4 days; cisplatin + conditioned medium group: a single intraperitoneal injection of 5 mg / kg body weight of cisplatin on the first day of treatment, and then continuous in...
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