title: The differentiation of human adipose-derived stem cells towards a urothelium-like phenotype in vitro and the dynamic temporal changes of related cytokines by both paracrine and autocrine signal regulation.
abstract: OBJECTIVE
  To investigate the differentiation ability of human adipose-derived stem cells (ASCs) towards urothelium-like cells in vitro and the dynamic changes of related cytokines and cytokine receptors in the culture medium.
  
  
  METHODS
  The ASCs were induced using both conditioned media (CM) and the transwell co-culture system with an immortalized urothelium cell line (SV-HUC-1,HUC) for 21 days. Protein and mRNA expression of the mature urothelium specific markers uroplakin-IA (UP-1A) and uroplakin-II (UP-II) were detected by immunofluorescence and quantitative real-time PCR, respectively. Array detection was used to screen 41 cytokines and receptors in the upper medium of urothelium, non-induced ASCs and urothelium-induced ASCs at three time points, early (12 hours), intermediate (7 days) and late (21 days).
  
  
  RESULTS
  After induction for 7 days, the ASCs grown in both CM and transwell co-culture system expressed uroplakin-IA (13.54±2.00%; 17.28±1.84%) and uroplakin-II (19.49±1.73%; 13.98±1.47%). After induction for 21 days, ASCs grown in co-culture had significantly increased expression of uroplakin-IA (48.03±1.25%; 49.57±2.85%) and uroplakin-II (45.38±2.50%; 46.58±1.95%). In the upper medium of urothelium, 28 cytokines and 8 cytokine receptors had significantly higher expression than the counterpart of non-induced ASCs. After 7 days induction, the expression of 22 cytokines and 8 cytokine receptors was significantly elevated in the upper medium of induced ASCs compared to non-induced ASCs. At the early and intermediate time points, ASCs secreted high levels of relative cytokines and soluble receptors, but their expressions decreased significantly at the late time point.
  
  
  CONCLUSIONS
  The adipose-derived stem cells have the potential to be differentiated into urothelium-like cells in vitro by both CM and transwell co-culture system with mature urothelium. Numerous cytokines and receptors were involved in the differentiation process with dynamic temporal changes by both paracrine and autocrine signal regulation. Further studies should be carried out to determine the detailed mechanism of cytokines and receptors and to enhance the urothelium differentiation efficiency of ASCs.
authors.affiliation: Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Ming-Xi
authors.initials: MX
authors.lastname: Xu
authors.affiliation: Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Mu-Jun
authors.initials: MJ
authors.lastname: Lu
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D019898:Autocrine Communication; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D048430:Cell Shape; D016207:Cytokines / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D019899:Paracrine Communication; D010641:Phenotype; D018121:Receptors, Cytokine / Q000378:metabolism; D012995:Solubility; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D019459:Urothelium / Q000166:cytology*
chemical_list: D015415:Biomarkers; D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; D018121:Receptors, Cytokine
references.citation: PLoS One. 2012;7(12):e52405
references.pmid: 23300662
references.citation: Tissue Eng. 2001 Apr;7(2):211-28
references.pmid: 11304456
references.citation: World J Urol. 2010 Aug;28(4):493-8
references.pmid: 20091038
references.citation: Biomaterials. 2009 Jul;30(19):3259-70
references.pmid: 19345408
references.citation: Stem Cells. 2012 Mar;30(3):481-90
references.pmid: 22213158
references.citation: J Sex Med. 2009 Apr;6(4):967-979
references.pmid: 19207272
references.citation: PLoS One. 2012;7(7):e40820
references.pmid: 22808268
references.citation: Proc Natl Acad Sci U S A. 2012 Feb 7;109(6):2090-5
references.pmid: 22308363
references.citation: Proc Natl Acad Sci U S A. 2006 Aug 8;103(32):12167-72
references.pmid: 16880387
references.citation: Cell Tissue Res. 2012 Jan 31;347(3):737-746
references.pmid: 22290635
references.citation: Methods Mol Biol. 2013;1001:65-80
references.pmid: 23494421
references.citation: PLoS One. 2013 Sep 12;8(9):e73266
references.pmid: 24069182
references.citation: PLoS One. 2010 May 19;5(5):e10706
references.pmid: 20502661
references.citation: Urology. 2010 Jun;75(6):1514.e7-14
references.pmid: 20394977
references.citation: Keio J Med. 2005 Sep;54(3):132-41
references.pmid: 16237275
references.citation: J Urol. 2008 Oct;180(4 Suppl):1778-83
references.pmid: 18721942
references.citation: Am J Physiol Lung Cell Mol Physiol. 2010 Jun;298(6):L735-43
references.pmid: 20190033
references.citation: Cytotechnology. 2011 Oct;63(5):531-9
references.pmid: 21915725
references.citation: Stem Cell Rev Rep. 2011 Mar;7(1):64-76
references.pmid: 20396979
references.citation: Tissue Eng Part A. 2010 May;16(5):1769-79
references.pmid: 20020816
references.citation: Biochem Biophys Res Commun. 2009 Dec 18;390(3):931-6
references.pmid: 19852942
references.citation: Biochem Biophys Res Commun. 2012 Nov 16;428(2):271-7
references.pmid: 23085045
references.citation: Urology. 2013 Feb;81(2):465.e15-22
references.pmid: 23374843
references.citation: Tissue Eng Part A. 2013 Jan;19(1-2):235-46
references.pmid: 22924692
references.citation: PLoS One. 2013 May 28;8(5):e63638
references.pmid: 23723991