title: Pathologic bladder microenvironment attenuates smooth muscle differentiation of skin derived precursor cells: implications for tissue regeneration.
abstract: Smooth muscle cell containing organs (bladder, heart, blood vessels) are damaged by a variety of pathological conditions necessitating surgery or organ replacement. Currently, regeneration of contractile tissues is hampered by lack of functional smooth muscle cells. Multipotent skin derived progenitor cells (SKPs) can easily be isolated from adult skin and can be differentiated in vitro into contractile smooth muscle cells by exposure to FBS. Here we demonstrate an inhibitory effect of a pathologic contractile organ microenvironment on smooth muscle cell differentiation of SKPs. In vivo, urinary bladder strain induces microenvironmental changes leading to de-differentiation of fully differentiated bladder smooth muscle cells. Co-culture of SKPs with organoids isolated from ex vivo stretched bladders or exposure of SKPs to diffusible factors released by stretched bladders (e.g. bFGF) suppresses expression of smooth muscle markers (alpha SMactin, calponin, myocardin, myosin heavy chain) as demonstrated by qPCR and immunofluorescent staining. Rapamycin, an inhibitor of mTOR signalling, previously observed to prevent bladder strain induced de-differentiation of fully differentiated smooth muscle cells in vitro, inhibits FBS-induced smooth muscle cell differentiation of undifferentiated SKPs. These results suggest that intended precursor cell differentiation may be paradoxically suppressed by the disease context for which regeneration may be required. Organ-specific microenvironment contexts, particularly prevailing disease, may play a significant role in modulating or attenuating an intended stem cell phenotypic fate, possibly explaining the variable and inefficient differentiation of stem cell constructs in in vivo settings. These observations must be considered in drafting any regeneration strategies.
authors.affiliation: Developmental and Stem Cell Biology, Research Institute, Toronto, Ontario, Canada.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Tolg
authors.forename: Alya
authors.initials: A
authors.lastname: Ahsan
authors.forename: Shaalee
authors.initials: S
authors.lastname: Dworski
authors.forename: Tyler
authors.initials: T
authors.lastname: Kirwan
authors.forename: Jeffery
authors.initials: J
authors.lastname: Yu
authors.forename: Karen
authors.initials: K
authors.lastname: Aitken
authors.forename: Darius Jehan
authors.initials: DJ
authors.lastname: Bägli
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects; D054337:Cell Dedifferentiation / Q000187:drug effects; D002454:Cell Differentiation; D060833:Cellular Microenvironment; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005260:Female; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D006801:Humans; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012038:Regeneration; D012867:Skin / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013314:Stress, Mechanical; D046509:Tissue Culture Techniques; D001743:Urinary Bladder / Q000378:metabolism* / Q000473:pathology*
chemical_list: D015415:Biomarkers; D017077:Culture Media, Conditioned; D005346:Fibroblast Growth Factors
references.citation: Cell Tissue Res. 2008 Sep;333(3):449-59
references.pmid: 18607632
references.citation: Adv Dent Res. 1998 Nov;12(2):12-26
references.pmid: 9972117
references.citation: World J Urol. 2010 Aug;28(4):493-8
references.pmid: 20091038
references.citation: J Urol. 2005 Oct;174(4 Pt 2):1699-703
references.pmid: 16148685
references.citation: Neurourol Urodyn. 1995;14(4):297-309
references.pmid: 7581466
references.citation: Urol Int. 2008;81(2):222-7
references.pmid: 18758225
references.citation: Cardiovasc Res. 2006 Apr 1;70(1):136-45
references.pmid: 16451796
references.citation: J Urol. 2004 Mar;171(3):1348-52
references.pmid: 14767346
references.citation: Arterioscler Thromb Vasc Biol. 2003 Mar 1;23(3):483-8
references.pmid: 12615694
references.citation: J Vasc Res. 2001 Mar-Apr;38(2):120-32
references.pmid: 11316948
references.citation: Lab Invest. 1999 Nov;79(11):1335-45
references.pmid: 10576204
references.citation: Cardiovasc Res. 2002 Sep;55(4):850-63
references.pmid: 12176134
references.citation: Philos Trans R Soc Lond B Biol Sci. 2008 Jan 12;363(1489):185-98
references.pmid: 17282990
references.citation: Methods Mol Biol. 2009;482:159-70
references.pmid: 19089355
references.citation: J Neurosci. 2007 Sep 5;27(36):9545-59
references.pmid: 17804616
references.citation: Urology. 2001 Feb;57(2):371-5
references.pmid: 11182367
references.citation: Nat Cell Biol. 2001 Sep;3(9):778-84
references.pmid: 11533656
references.citation: J Biomed Mater Res A. 2012 Jan;100(1):72-83
references.pmid: 21972045
references.citation: J Mol Med (Berl). 1999 Feb;77(2):294-8
references.pmid: 10023782
references.citation: Arthritis Rheum. 1999 Jun;42(6):1140-6
references.pmid: 10366106
references.citation: J Smooth Muscle Res. 2007 Apr;43(2):55-72
references.pmid: 17598958
references.citation: J Urol. 1999 Sep;162(3 Pt 1):832-40
references.pmid: 10458391
references.citation: Methods Mol Biol. 2012;821:361-72
references.pmid: 22125078
references.citation: J Urol. 2008 Oct;180(4 Suppl):1778-83
references.pmid: 18721942
references.citation: World J Urol. 2008 Aug;26(4):341-9
references.pmid: 18654786
references.citation: Can J Neurol Sci. 2010 Sep;37 Suppl 2:S34-41
references.pmid: 21246933
references.citation: J Urol. 2009 Oct;182(4 Suppl):1898-905
references.pmid: 19695585
references.citation: Physiol Genomics. 2004 Dec 15;20(1):36-44
references.pmid: 15467014
references.citation: Stem Cells Dev. 2009 Jul-Aug;18(6):893-906
references.pmid: 18834279
references.citation: J Urol. 1998 Mar;159(3):998-1005
references.pmid: 9474216
references.citation: Am J Pathol. 2010 Jan;176(1):304-19
references.pmid: 20019183
references.citation: Cell Struct Funct. 1997 Feb;22(1):65-72
references.pmid: 9113392
references.citation: Nat Rev Urol. 2009 Nov;6(11):596-611
references.pmid: 19890339
references.citation: Arterioscler Thromb Vasc Biol. 2009 Feb;29(2):232-8
references.pmid: 19074484
references.citation: J Urol. 2003 Nov;170(5):2072-6
references.pmid: 14532856
references.citation: Mol Cell. 2006 Apr 21;22(2):159-68
references.pmid: 16603397
references.citation: Am J Pathol. 2006 Aug;169(2):459-70
references.pmid: 16877348
references.citation: Nat Protoc. 2006;1(6):2803-12
references.pmid: 17406538
references.citation: Anat Rec A Discov Mol Cell Evol Biol. 2003 Oct;274(2):883-6
references.pmid: 12973712
references.citation: Nat Cell Biol. 2004 Nov;6(11):1082-93
references.pmid: 15517002
references.citation: Vitam Horm. 2011;87:127-41
references.pmid: 22127241
references.citation: Lancet. 2006 Apr 15;367(9518):1241-6
references.pmid: 16631879
references.citation: Stem Cells. 2005 Jun-Jul;23(6):727-37
references.pmid: 15917469
references.citation: Cell Stem Cell. 2009 Dec 4;5(6):610-23
references.pmid: 19951689
references.citation: Adv Dent Res. 1998 Nov;12(2):114-8
references.pmid: 9972133
references.citation: Proc Natl Acad Sci U S A. 2009 May 12;106(19):7840-5
references.pmid: 19416884
references.citation: J Invest Dermatol. 2010 May;130(5):1202-4
references.pmid: 20393474
references.citation: Am J Physiol Renal Physiol. 2007 Oct;293(4):F1007-17
references.pmid: 17634401
references.citation: In Vitro Cell Dev Biol Anim. 2006 May-Jun;42(5-6):124-33
references.pmid: 16848631
references.citation: Urology. 2010 Mar;75(3):664-70
references.pmid: 19963239
references.citation: Mol Cell Biol. 2009 Mar;29(6):1411-20
references.pmid: 19114562
references.citation: Arterioscler Thromb Vasc Biol. 2011 Dec;31(12):2938-48
references.pmid: 21852558
references.citation: World J Stem Cells. 2010 Feb 26;2(1):1-4
references.pmid: 21607109