Improvement of a procedure for preparing single smooth muscle cells from guinea pig tenia coli by purified collagenase and papain.
Procedures for isolating single smooth muscle cells from tenia coli of guinea pigs have been developed. The cells were isolated with a combination of highly purified collagenase and a small amount of strong peptidase, papain, in the presence of Ca. This combination resulted in approximately a 100-fold yield of the single cells as compared with the ordinary method in which only collagenase preparations contaminated with various peptidases were used. More than 95% of the single cells were viable when examined by the trypan blue exclusion technique. In addition, this combination greatly increased the responsiveness to agonists such as carbachol (CCh). A concentration of CCh for the half maximum response for this preparation was about one five-hundredth that of cells prepared by the ordinary method. This value was approximately one-quarter that of intact tissue. This preparation can be used to investigate the physiology, pharmacology, and biochemistry of smooth muscle.
Maruyama
I
I
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
Yoshida
C
C
Nakayama
M
M
Hasegawa
T
T
Momose
K
K
D000818:Animals; D001285:Atropine / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D003106:Colon / Q000166:cytology; D006168:Guinea Pigs; D008722:Methods; D003012:Microbial Collagenase / Q000494:pharmacology*; D009130:Muscle, Smooth / Q000166:cytology*; D010206:Papain / Q000494:pharmacology*
D001285:Atropine; D002217:Carbachol; D010206:Papain; D003012:Microbial Collagenase