Studies on isolated smooth muscle cells. IX. Application of papain for isolation of single smooth muscle cells from guinea-pig taenia coli.
To prepare single smooth muscle cells from the taenia coli of guinea pig, the application of papain to the enzymatic solution was examined under two conditions: 1) the isolation in a modified Tyrode solution (containing 0.18 mM Ca2+: 0.18 mM Ca2+-Tyrode solution) and 2) the isolation in a high-K+ Tyrode solution (Na+ was replaced by K+, and Ca2+ was not added: high-K+ Tyrode solution). The presence of papain during collagenase digestion reduced contamination of broken cells and cell debris. In the case of the high-K+ Tyrode solution, papain increased the yield of single cells significantly. The cells were contracted in a dose-dependent manner by Ca2+ in the high-K+ Tyrode solution and by carbachol in 0.18 mM Ca2+-Tyrode solution; furthermore, the contractions were antagonized by verapamil and atropine, respectively. Treatment with papain did not affect cell sensitivity to the stimulants. Therefore, our results suggest that the addition of papain is useful for the isolation of single cells to investigate the physiological and pharmacological characteristics of smooth muscle.
Hasegawa
M
M
University of Showa, School of Pharmaceutical Sciences, Tokyo, Japan.
Kobayashi
M
M
Oyamada
H
H
Kamishima
T
T
Yoshida
C
C
Ohata
H
H
Maruyama
I
I
Momose
K
K
Gomi
Y
Y
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D002452:Cell Count; D002469:Cell Separation / Q000379:methods; D005260:Female; D006168:Guinea Pigs; D008297:Male; D009130:Muscle, Smooth / Q000166:cytology*; D010206:Papain / Q000494:pharmacology*
D002217:Carbachol; D010206:Papain; D002118:Calcium