Momose K, Gomi Y
Nihon Heikatsukin Gakkai Zasshi. 1980 Mar;16(1):29-36. doi: 10.1540/jsmr1965.16.29.
Dispersion procedure of smooth muscle cells from collagenase-treated minces of guinea pig taenia coli and their response to acetylcholine were examined in order to apply the single smooth muscle cells to investigation of contraction mechanisms of smooth muscle. Pipetting the mince suspension was convenient procedure for the dispersion of single cells. For good yield of acetylcholine-sensitive cells, very mild pipetting was required although yield of single cells was lower. Under the condition, 10(3)-10(4) cells were obtained from 3-4 cm of taenia coli strip and about a half of the cells were responsive to acetylcholine. The cells became 40% of the original length by administration of 10 mM acetylcholine. Once the cells were dispersed and suspended in the physiological salt solution the cells lost the sensitivity to acetylcholine within 90 minutes while very mild pipetting of the mince suspension provided acetylcholine-sensitive cells over 3 hours. These single cells were applicable to investigation of contraction mechanisms of smooth muscle in addition to whole tissue as used so far. It was also suggested in the experiment that acetylcholine-sensitive site of cells might be damaged by exposure to physiological salt solution while it was well preserved in the tissue even though the tissue was digested by collagenase.
为了将单个平滑肌细胞应用于平滑肌收缩机制的研究，对从经胶原酶处理的豚鼠结肠带切碎组织中分散平滑肌细胞的方法及其对乙酰胆碱的反应进行了研究。用移液管吸取切碎组织悬液是分散单个细胞的简便方法。为了获得高产率的对乙酰胆碱敏感的细胞，尽管单个细胞的产率较低，但需要非常轻柔地移液。在这种条件下，从3 - 4厘米的结肠带条中可获得10³ - 10⁴个细胞，且约一半的细胞对乙酰胆碱有反应。给予10 mM乙酰胆碱后，细胞长度变为原来的40%。一旦细胞被分散并悬浮于生理盐溶液中，细胞在90分钟内就会失去对乙酰胆碱的敏感性，而非常轻柔地移液处理切碎组织悬液可使细胞在3小时以上保持对乙酰胆碱的敏感性。这些单个细胞除了可像迄今为止使用的整个组织一样用于平滑肌收缩机制的研究外，实验还表明，细胞对乙酰胆碱敏感的部位可能会因暴露于生理盐溶液而受损，而在组织中即使组织被胶原酶消化，该部位仍能得到很好的保存。
