A method for isolation of adult cardiac mouse myocytes for studies
of contraction and micro-fluorimetry.
Wolska, Beata M., and R. John Solaro.
Department of Physiology and Biophysics, University of Illinois at
Chicago, College of Medicine, Chicago, IL 60612-7342
APStracts 3:0047H, 1996.
We describe techniques for the isolation of Ca2+-tolerant myocytes
from mouse (2-6 months old) ventricle for measurements of mechanics
of contraction and micro-fluorimetry. Our approach involved special
modifications of existing methods that had been developed for other
species, but were not successful when applied to the mouse heart.
Important features of the method are: i) a requirement for careful
timing (&LT5 min) of perfusion with nominally Ca2+-free
solution, ii) perfusion with a solution containing a specially
selected batch of collagenase in the presence of a low concentration
of Ca2+, iii) meticulous attention to water quality. Using this
method, we could consistently isolate durable, Ca-tolerant myocytes
from adult mouse hearts with a yeild of around 50%. With slight
modifications, the method should enable other investigators to
isolate mouse cardiomyocytes for their specific experimental
applications.
Received 6 September 1995; accepted in final form 15 January
1996.
APS Manuscript Number H842-5.
Article publication pending Am. J. Physiol. (Heart Circ. Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 8 February 96