| Objective : To explore the therapeutic effects of mouse adipose tissue-derived stromal stem cells transplantation on injured endometrium in vivo,which provides experimental and theoretical basis for the treatment aiming at thin endometrium,so as to improve the pregnancy rate.Methods:1 The establishment of injured endometrium within mice: KM female mice,aging 6-8 weeks,were intraperitoneally injected with PMSG in order to synchronize the estrus cycles.After 48 hours,they were divided into 3 groups at random: experimental group were perfused 95%ethanol(injured for 5 minutes),control group were perfused DPBS(impacted for 5 minutes),blank group(none done).In 2 hours?the first and second estrum(determined by the vaginal fluid smear)after injury,endometrium thickness and the amount of glands of endometrium were observed through HE stain of tissue sections;Fibrosis was recorded via Masson stain.2 The frozen GFP-ADSCs were thawed and cultured.48 hours after cultivating,the cells were digested with trypsin,which were seeded into cultured fluid mixed with 3D hydrogel afterwards.3 The transplantation of GFP-ADSCs on injured endometrium: Mice with endometrium damage were divided into 3 group.4 hours after molding,the treatment group was perfused the cell suspension containing GFP-ADSCs and 3D hydrogel;control group is consist of two groups: Group 3D hydrogel was perfused the miscible fluid of 3D hydrogel and culture fluid,marked 3D hydrogel as control 1;Group DPBS was perfused DPBS as control group 2;the blank group has nothing to deal with.In the first and second estrum after transplantation,endometrium thickness and the amount of glands of endometrium were observed through HE stain.Fibrosis was recorded via Masson stain.In addition,green fluorescence light was observed on tissue sections derived from treatment group by means of fluorescence microscope.Results:1 In 2 hours and the first estrum after the operation,the endometrium thickness of experimental groups were smaller,which was significantly different from control and blank group(p<0.05),but the other groups were no difference as for each other.However,in the second estrum,the observed values of the thickness of experimental group was still smaller than the rest two groups,which is not reflected in statistics(p>0.05).In 2 hours,the first and second estrum,the glands of experimental group were smaller than the other two groups,whose date significantly varies(p<0.05),but the amounts of control and blank groups are similar(p>0.05).In addition,the area of fibrosis was obviously wider than control and blank group in 2 hours,the first and second estrum(p<0.05),but the other two groups made no difference(p>0.05).2 In the process of culturing,we observed that the cells grew adherent to the wall taken on appearances as long spindles.3 In the first estrum after the procedure of transplantation,the average thickness of endometrium of treatment group was 424.51 um that is close to the number of blank group(394.63?m)(p>0.05),but it reflected difference in the comparison to the other two control groups(279.77?m,265.91?m)(p<0.05).In the second estrum,the thickness of treatment group was 642.21?m,which stood for the wide difference from the other three groups(p<0.05).The amount of glands of treatment group were larger than control group(p<0.05),but it has no difference from blank group(p>0.05).The degree of fibrosis was significantly reduced compared to control group(P<0.05),but it made no difference from blank group,In each stage,the two control groups had no difference in terms of three injury indicators in statistics.Moreover,we found there was no green fluorescence light observed on the tissue sections in the first or second estrum after the treatment.Conclusions: The mouse model of injured endometrium could be established through the95% alcohol perfused into uterus.The transplantation of GFP-ADSCs into uterus could promote repairing the endometrium. |
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