Isolation And Purification Of Endometrial Mesenchymal Stem Cells And Its Clinical Observation In The Treatment Of AS Syndrome

Asherman syndrome(AS),also known as intrauterine adhesions(IUAS),refers to the destruction of the endometrial basal layer,this can cause partial or total adhesion of the uterine cavity,gynecological diseases such as periodic pelvic pain,abnormal menstruation,repeated abortion,infertility and other symptoms.And the destruction of the endometrial basal layer usually caused by intrauterine surgery,intrauterine infection,radiation and other factors.Recent studies have suggested that there may be stem cells/progenitor cells in the uterine lining of the uterus,and the high occurrence and recurrence rate of intrauterine adhesions may be related to the reduction,loss and dysfunction of endometrial stem cells.so it is proposed to use stem cell transplantation as a new methods to treat intrauterine adhesions and to prevent recurrence.Our study use the intrauterine perfusion of endometrial mesenchymal stem cells to treat moderate and severe AS syndrome.In this study,we sought a appropriate treatment for AS syndrome by comparing the changes in endometrial thickness,subendocardial blood flow,endometrial receptivity,and embryo implantation rate between the two cell treatments.The main contents include: the first part,the seperation,cultivation and identification of endometrial mesenchymal stem cells.the second part,randomly divided 45 cases of moderate or severe intrauterine adhesions into experimental group A,experimental B and control group C,15 cases in each group.All patients underwent hysteroscopic intrauterine adhesions(TCRA)after admission,experimental group A was treated with endometrial mesenchymal stem cells alone,experimental group B was treated with endometrial mesenchymal stem cells and sodium hyaluronate gel,control group C only treated with sodium hyaluronate gel.All patients underwent embryo transfer in the fourth month after surgery.Postoperative gynecological color doppler ultrasound and hysteroscopy were performed in the outpatient department.The endometrial thickness,American Fertility Association(AFS)score,endometrial blood flow distribution type,PI,S/D and pregnancy status were recorded.All case had on infection,endometriosis,pulmonary embolism and other complications after stem cell transplantation.Results:(1)The eMSC cells isolated in this experiment have uniform morphology,stable phenotype and strong proliferation ability in vitro.The results of flow cytometry showed that they were consistent with the surface markers of mesenchymal stem cells,and the positive rate was higher than 98%.(2)Endometrial mesenchymal stem cells can survive and grow well in sodium hyaluronate gel.(3)when the age is under 35,treatment with eMSCs transplantation alone and treatment with the combination of eMSCs and sodium hyaluronate gel can effectively promote the regeneration and repair of endometrial.When the age is over or equal to 35,combined use of eMSCs and sodium hyaluronate gel for treatment can promote the proliferation of the endometrial.(4)when the age is under 35,the treatment of eMSCs combined with sodium hyaluronate gel can effectively promote subendometrial angiogenesis and improve endometrial receptivity.When the age is over or equal to 35,the treatment of stem cells alone and combined use of stem cells and sodium hyaluronate gel can improve the subendometrial blood flow perfusion ability and improve endometrial receptivity.(5)when age is under 35,the combined treatment of stem cells and sodium hyaluronate gel can significantly improve the postoperative pregnancy rate of patients with AS syndrome.Conclusion: The results of this experiment indicate that autologous endometrial mesenchymal stem cells combined with sodium hyaluronate gel can promote the regeneration and repair of impaired endometrium,improve endometrial receptivity,restore endometrial function,and finally achieve successful pregnancy.These indicate that autologous endometrial mesenchymal stem cells combined with sodium hyaluronate gel are feasible for the treatment of AS syndrome.