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Objective To explore a simple and efficient method for isolating and culturing mouse primordial germ cells (mPGCs) in vitro in order to provide the sufficient mPGCs sources of mouse model .Methods The 12 .5 dpc mouse embryonic gen-ital ridge were isolated in vitro and cut into pieces ,then the organization to adherent culture was performed in a-MEM medium con-taining fetal calf serum and insulin-transferrin-selenium (ITS) and follicle stimulating hormone(FSH) and recombinant endothelial growth factor(rEGF) .The inverted microscope was used to observe the cellular morphology .Flow cytometry was used to identify the stage specific embryonic antigens of cultured cells .Reverse transcription-polymerase chain reaction(RT-PCR) and immunofluo-rescence were used to identify the pluripotency gene Oct4 expressions of stem cells and specific genes in the meiosis phaseⅠ .Results The in vitro isolated and cultured mPGCs showed good cell morphology ,extremely strong proliferation capacity and the positive expression of SSEA-1 and negative expression of SSEA-4 ,Oct4 ,meanwhile the expressions of Stra8 ,Vasa ,Scp3 ,Zp3 were detected to be positive .Conclusion Using this culture method can culture the high purity of mPGCs with the excellent stem cell properties and extremely strong proliferative ability ,moreover which makes the cells entering the meiosis stage .
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Objective To study the desired therapeutic effect of vaginoplasty using sigmoid colon or peritoneum for entire vaginal atre-sia.Methods From May 2015 to October 2015,8 patients with complete vaginal atresia in our hospital performed vaginoplasty using sigmoid colon or peritoneum successfully.The operation methods,prognosis and therapeutic effect and quality of sexual life of two groups were com-pared and analyzed to investigate a perfect operative method and ideal nursing.Results Laparoscopic vaginoplasty using the peritoneum compared with that using sigmoid colon has advantages of shorter time of operation,less bleeding,less trauma and quicker recovery.However, artificial vagina using sigmoid colon was much more similar to a natural vagina in morphological and physiological characteristics,merely ac-companied by excessive mucus discharge during the postoperative period.At lower risk of vaginal adhesion and stenosis,laparoscopically as-sisted vaginoplasty using sigmoid colon graft also puts high technical requirements.Two patients normally developed uterus menstruate regular-ly.Four patients without uterus and 2 patients with rudimentary uterus experienced good sexual intercourse after vaginoplasty.Conclusion Patients with complete colpatresia who have normal womb should creat a patent and functional vagina until adolescence to restore its physio-logical and reproductive function.Anatomical reconstructive treatment of vaginal agenesis for patients with absent or hypoplastic uterus can be postponed till the late teens or in the adult.Both laparoscopic sigmoid vaginoplasty and peritoneal vaginoplasty achieve a minimal-invasive, cosmetic,natural lubricous, smooth artificial vagina for patient’ s sex life satisfactory.