Zuurveld J G, Veerkamp J H, Wirtz P
Muscle Nerve. 1985 Nov-Dec;8(9):750-9. doi: 10.1002/mus.880080903.
Myofiber suspensions with 80% viability (morphological intactness) were prepared from rat m. flexor digitorum brevis by a 6-hour collagenase treatment. The viable fibers had intact sarcolemmas and generally showed continuous basal laminae. The intracellular compartments exhibited normal muscular morphology. The suspended myofibers showed good tolerance against extracellular calcium (up to 30 mM), but were susceptible to 15 mM EGTA. Calcium ionophore A23187 induced contraction, with loss of viability in both calcium-containing and nominally calcium-free Krebs-Ringer medium. EGTA prevented the ionophore effect, but the muscle cells collapsed as soon as calcium was replenished. A calcium paradox, as observed with perfused heart, could not be induced in isolated skeletal myocytes. This suspended cell system should provide a useful tool for investigating relationships between metabolic parameters and muscle physiology. The extracellular environment can be manipulated easily, and cellular responses of individual skeletal muscle fibers can be measured and visualized.
通过6小时的胶原酶处理，从大鼠趾短屈肌制备出活力为80%（形态完整性）的肌纤维悬液。存活的纤维具有完整的肌膜，并且通常显示出连续的基膜。细胞内区室呈现出正常的肌肉形态。悬浮的肌纤维对细胞外钙（高达30 mM）表现出良好的耐受性，但对15 mM的乙二醇双四乙酸（EGTA）敏感。钙离子载体A23187诱导收缩，在含钙和名义上无钙的 Krebs-Ringer 培养基中均导致活力丧失。EGTA 可防止离子载体效应，但一旦补充钙，肌肉细胞就会立即塌陷。在分离的骨骼肌细胞中无法诱导出像灌注心脏中观察到的钙反常现象。这种悬浮细胞系统应为研究代谢参数与肌肉生理学之间的关系提供一个有用的工具。细胞外环境可以轻松操控，并且可以测量和观察单个骨骼肌纤维的细胞反应。
