title: How to isolate urothelial cells? Comparison of four different methods and literature review.
abstract: The aim of this study is to present the comparison of four different methods for urothelial cell isolation and culture and compare them to methods cited in the literature. Four different techniques were examined for urothelium isolation from rat bladders. Isolation effectiveness was calculated using trypan blue assay. Confirmation of isolated cell phenotype and comparison with native bladder tissue was confirmed using immunohistochemical (IHC), immunocytochemical (ICC) and immunofluorescence (IF) analysis. The method with bladder inversion and collagenase P digestion resulted in the highest number of isolated cells. These cells showed positive expression of cytokeratin 7, 8, 18, α6-integrin and p63. Our results and the literature review showed that the best method for urothelium bladder isolation is dissection of the epithelium layer from other bladder parts and digestion of mechanically prepared tissue in a collagenase solution.
authors.affiliation: Department of Tissue Engineering, Ludwik Rydygier Collegium Medicum in Bydgoszcz Nicolaus Copernicus University in Toruń, Karlowicza Str. 24, 85-092, Bydgoszcz, Poland, tomaszkloskowski@op.pl.
authors.forename: T
authors.initials: T
authors.lastname: Kloskowski
authors.forename: M
authors.initials: M
authors.lastname: Uzarska
authors.forename: N
authors.initials: N
authors.lastname: Gurtowska
authors.forename: J
authors.initials: J
authors.lastname: Olkowska
authors.forename: R
authors.initials: R
authors.lastname: Joachimiak
authors.forename: A
authors.initials: A
authors.lastname: Bajek
authors.forename: M
authors.initials: M
authors.lastname: Gagat
authors.forename: A
authors.initials: A
authors.lastname: Grzanka
authors.forename: M
authors.initials: M
authors.lastname: Bodnar
authors.forename: A
authors.initials: A
authors.lastname: Marszałek
authors.forename: T
authors.initials: T
authors.lastname: Drewa
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases / Q000378:metabolism; D039503:Integrin alpha6 / Q000378:metabolism; D053538:Keratin-18 / Q000378:metabolism; D053552:Keratin-7 / Q000378:metabolism; D053533:Keratin-8 / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D012038:Regeneration; D012996:Solutions; D001743:Urinary Bladder / Q000166:cytology*; D019459:Urothelium / Q000166:cytology* / Q000378:metabolism / Q000502:physiology
chemical_list: D039503:Integrin alpha6; D053538:Keratin-18; D053552:Keratin-7; D053533:Keratin-8; D012996:Solutions; D017364:Collagenases