The Role Of Adipose Mesenchymal Stem Cells In The Treatment Of Premature Ovarian Failure Model Mice

Objective:To simulate the symptoms of clinical premature ovarian failure,establish an animal model of premature ovarian failure,and explore the biological characteristics of adipose mesenchymal stem cells and their role in the treatment of premature ovarian failure model rats.In order to provide theoretical basis for clinical adipose mesenchymal stem cells in the treatment of patients with premature ovarian failure.Materials and methods:The subcutaneous fat in the bilateral groin of normal rats was isolated and cultured and subcultured in vitro.The cells after the third generation were selected for all experiments.Osteogenic and adipogenic differentiation of adipose mesenchymal stem cells were induced,and the morphological changes of adipose mesenchymal stem cells were observed by microscope.The phenotype of the cells was identified by Novo Cyte flow cytometry.The proliferation inhibition rate of cells was detected by CCK8 after24,48,72,96,120,144,168 hours.The rat model of POF was established by intraperitoneal injection of cisplatin.The rats were randomly divided into two groups:model group（Model）and ADMSCs treatment group,treatment group injected adipose mesenchymal stem cells through tail vein,model group injected normal saline through tail vein.The changes of body weight,serum estradiol and other hormones,ovarian tissue and pathological sections of rats in each group were observed.Results:1.Adipose mesenchymal stem cells are adherent growth cells,the initial cell morphology is pleomorphic,after purification,they are uniform and long fusiform,and when they are exuberant,they are vortex-like and fish-like.2.Adipose mesenchymal stem cells could differentiate into osteogenesis and adipogenesis.On the 21st day of osteogenic differentiation,the extracellular matrix produced deposited calcium nodules;14 days after adipogenic differentiation,a large number of lipid vacuoles of different sizes and good refraction could be seen in the cytoplasm.3.The positive expression rates of ADMSCs high expression CD44and CD90,were 75.85%and 59.71%respectively,the low expression rate of CD45 was low,and the negative expression rate was 39.12%.4.Adipose mesenchymal stem cells grew slowly from day 1 to day 2,grew rapidly from day 3,and grew logarithmically from day 6 to day 7.From day 6 to day 7,the growth rate entered a plateau and stopped growing,and the whole growth curve was S-shaped.5.Compared with the normal group,the body weight of the model group decreased significantly,and the serum hormone level of the model group was consistent with the clinical characteristics of premature ovarian failure,the levels of FSH and LH increased and the level of E₂decreased,which was statistically significant.According to the pathological sections of the ovary,the number of follicles at all levels in the model group was significantly lower than that in the normal group,and there were more atretic follicles.6.After treatment,the body weight of rats in the treatment group showed a trend of slow increase,but the body weight of the model group did not increase significantly,and there was no statistical significance between the two groups.In the serum of rats in the treatment group,the levels of E₂increased and the levels of FSH and LH decreased,so there was statistical significance,while for the normal group,the concentration of LH and FSH was higher and the level of E₂was lower,which was not statistically significant.From the results of ovarian pathological section,compared with the model group,the number of locked follicles in the treatment group decreased and the number of follicles at all levels increased,but there was no significant difference in the number and morphology of follicles between the treatment group and the normal group.Conclusion:1.POF model can be successfully established by intraperitoneal injection of cisplatin.2.ADMSCs can grow and reproduce in vitro,and has strong ability of growth,proliferation and multi-directional differentiation.3.ADMSCs can improve the serum E₂level and ovarian function of POF model rats,and has a certain effect in the treatment of premature ovarian failure.