Stem cell exosome composite preparation for improving premature ovarian failure and application of composite preparation
A compound preparation, the technology of premature ovarian failure, applied in the field of biology, can solve the problems of patients' psychological and physical distress, inability to achieve complete cure, repeated illness, etc., to improve the ovarian microenvironment, improve ovarian function, and achieve good curative effect. Effect
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Embodiment 1
[0035] Example 1: Extraction of exosomes from human endometrial stem cells
[0036] 1. Human endometrial stem cell culture: P5 generation human endometrial stem cells are selected, and the ratio is 2×10 5 / cm 2 Density inoculated at 75cm 2 Cell culture flask, and add DMEM / F12 medium containing 10% fetal bovine serum, at 37℃, 5% CO 2 Cultivate in an incubator for 2-3 days. When it grows to a density of 80%-85%, discard the original medium, and gently wash the cell surface twice with PBS phosphate buffer to remove cell surface debris and residual serum; The PBS solution was discarded, and 15 mL serum-free DMEM / F12 medium was added to each bottle to continue culturing for 72 hours.
[0037] 2. Collection of cell culture solution: After culturing endometrial stem cells in serum-free medium for about 48-72 hours, collect the cell culture solution aseptically, filter and sterilize it with a 0.22μm disposable filter, and store in aliquots at -20°C.
[0038] 3. Extraction of exosomes from th...
Embodiment 2
[0041] Example 2: Stem cell exosomes compound preparation improves premature ovarian failure in rats
[0042] 1. Preparation of experimental animals: Screen 40 2-month-old wistar female rats with normal estrus cycle, weighing 180±20g, after adaptive feeding for 7 days, weigh again, and randomly assign 10 rats as model control group , And the remaining 30 are modeled.
[0043] 2. Establishment of a rat model of premature ovarian failure: give rats a suspension of Tripterygium wilfordii polyglycoside tablets 50mg / kg daily for 14 days, take a smear of vaginal secretions with a cotton swab at 8:00 every day, and observe after Pap staining Changes in vaginal exfoliated cells. After the gavage, blood was collected from the tail vein of the rats to detect the levels of estradiol and follicle stimulating hormone in the blood and compare them with the modeled control group. When the continuous estrus interval was observed for 10 consecutive days, the serum estradiol level was significantl...
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