Maruyama I, Yoshida C, Nakayama M, Hasegawa T, Momose K
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
J Pharmacol Methods. 1988 Apr;19(2):155-64. doi: 10.1016/0160-5402(88)90036-8.
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.
已开发出从豚鼠结肠带分离单个平滑肌细胞的方法。在钙离子存在的情况下，使用高纯度胶原酶和少量强蛋白酶木瓜蛋白酶的组合来分离细胞。与仅使用被各种肽酶污染的胶原酶制剂的普通方法相比，这种组合使单个细胞的产量提高了约100倍。用台盼蓝排斥技术检测时，超过95%的单个细胞具有活力。此外，这种组合极大地增强了对诸如卡巴胆碱（CCh）等激动剂的反应性。该制剂产生半数最大反应的CCh浓度约为用普通方法制备的细胞的五百分之一。该值约为完整组织的四分之一。这种制剂可用于研究平滑肌的生理学、药理学和生物化学。
