An in vitro model for videoimaging of human bladder smooth muscle cell contractions.
Knowledge regarding human bladder smooth muscle cell (SMC) physiology is very limited. Only a few specific medical therapies for bladder disorders have therefore been established. The objective of this study was to develop a model for videomicroscopy of bladder SMC contractions. Cells were isolated from human cystoprostatectomy specimens and cultured in a modified EMEM medium. These cells were identified as SMCs by means of immunohistochemistry. For videomicroscopy, the culture flasks were coated with a viscous agent to allow cell contraction. Contractions were visualized by means of a cell culture microscope with a time-lapse videosystem. For cholinergic stimulation of the cells, acetylcholine, in concentrations ranging from 100 microM to 10 mM, was applied. The percentage of contracting cells within the observation field was evaluated for quantitative analysis. In control experiments without contractile stimulant 6% of the cells were observed to contract. Stimulation with acetylcholine induced a significant dose-dependent increase to 47% in contracting cells. These results demonstrated that videomicroscopy is an appropriate tool to investigate the contraction mechanisms of bladder SMCs. This model offers the possibility of studying drug effects on the human detrusor in vitro.
Corvin
S
S
Department of Urology, Ludwig-Maximilians-Universität, Munich, Germany. Stefan.Corvin@uro.med.uni-muenchen.de
Bösch
S T
ST
Maneschg
C
C
Bartsch
G
G
Klocker
H
H
D000109:Acetylcholine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008297:Male; D018715:Microscopy, Video; D009119:Muscle Contraction; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D014407:Tumor Cells, Cultured; D001743:Urinary Bladder / Q000166:cytology / Q000187:drug effects / Q000502:physiology*
D000109:Acetylcholine