title: Long-term culture and significant expansion of human Sertoli cells whilst maintaining stable global phenotype and AKT and SMAD1/5 activation.
abstract: BACKGROUND
  Sertoli cells play key roles in regulating spermatogenesis and testis development by providing structural and nutritional supports. Recent studies demonstrate that Sertoli cells can be converted into functional neural stem cells. Adult Sertoli cells have previously been considered the terminally differentiated cells with a fixed and unmodifiable population after puberty. However, this concept has been challenged. Since the number of adult human Sertoli cells is limited, it is essential to culture these cells for a long period and expand them to obtain sufficient cells for their basic research and clinic applications. Nevertheless, the studies on human Sertoli cells are restricted, because it is difficult to get access to human testis tissues.
  
  
  RESULTS
  Here we isolated adult human Sertoli cells with a high purity and viability from obstructive azoospermia patients with normal spermatogenesis. Adult human Sertoli cells were cultured with DMEM/F12 and fetal bovine serum for 2 months, and they could be expanded with a 59,049-fold increase of cell numbers. Morphology, phenotypic characteristics, and the signaling pathways of adult human Sertoli cells from different passages were compared. Significantly, adult human Sertoli cells assumed similar morphological features, stable global gene expression profiles and numerous proteins, and activation of AKT and SMAD1/5 during long-period culture.
  
  
  CONCLUSIONS
  This study demonstrates that adult human Sertoli cells can be cultured for a long period and expanded with remarkable increase of cell numbers whilst maintaining their primary morphology, phenotype and signaling pathways. This study could provide adequate human Sertoli cells for reproductive and regenerative medicine.
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China. kaoyanyong1@126.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China. hygeia@sjtu.edu.cn.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Hai
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China. yaochencheng@126.com.
authors.forename: Chencheng
authors.initials: C
authors.lastname: Yao
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China. zhengchenseu@126.com.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China. houjingmei@qq.com.
authors.forename: Jingmei
authors.initials: J
authors.lastname: Hou
authors.affiliation: Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Andrology, Shanghai Human Sperm Bank, 145 Shangdong Road, Shanghai, 200001, China. lizhengboshi@163.com.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China. zupinghe@sjtu.edu.cn.|Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Andrology, Shanghai Human Sperm Bank, 145 Shangdong Road, Shanghai, 200001, China. zupinghe@sjtu.edu.cn.|Shanghai Key Laboratory of Assisted Reproduction and Reproductive Genetics, Shanghai, 200127, China. zupinghe@sjtu.edu.cn.|Shanghai Key Laboratory of Reproductive Medicine, Shanghai, 200025, China. zupinghe@sjtu.edu.cn.
authors.forename: Zuping
authors.initials: Z
authors.lastname: He
mesh_terms: D000328:Adult; D000818:Animals; D053713:Azoospermia / Q000378:metabolism / Q000473:pathology; D002417:Cattle; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D012708:Sertoli Cells / Q000378:metabolism* / Q000473:pathology; D051898:Smad1 Protein / Q000378:metabolism*; D051902:Smad5 Protein / Q000378:metabolism*; D013997:Time Factors
chemical_list: C497399:SMAD1 protein, human; C497797:SMAD5 protein, human; D051898:Smad1 Protein; D051902:Smad5 Protein; D051057:Proto-Oncogene Proteins c-akt