Mangos J A, McSherry N R, Butcher F, Irwin K, Barber T
Am J Physiol. 1975 Sep;229(3):553-9. doi: 10.1152/ajplegacy.1975.229.3.553.
Morphologically and functionally intact acinar cells have been obtained from the rat parotid gland through enzymatic dispersion with pure collagenase, hyaluronidase, and trypsin as well as mild mechanical forces. Cell yields of 30-50% of the original tissue weight with over 95% acinar cells were accomplished. The cells in suspension assumed a more or less spherical shape but the intracellular polarity of organelle distribution was maintained. The cells in suspension at 37 degrees C maintained stable monovalent cationic composition but lost potassium and gained sodium rapidly upon exposure to ouabain, 10(-5) M. The intracellular amylase concentration and the patterns of secretion of amylase and of synthesis of cyclic AMP by the cells in response to adrenergic stimulation with epinephrine or isoproterenol were comparable to those of the intact gland in situ. In addition, the cells showed good O2 consumption and maintained it constant for periods up to 8 h. These cells could be used as experimental tools for in vitro studies of receptor physiology and biochemistry, cell membrane function, cellular secretory mechanisms, and other parameters of exocrine gland cell physiology.
通过用纯胶原酶、透明质酸酶和胰蛋白酶进行酶解分散以及施加温和的机械力,已从大鼠腮腺中获得了形态和功能完整的腺泡细胞。实现了细胞产量达到原始组织重量的30% - 50%,且腺泡细胞超过95%。悬浮细胞大致呈球形,但细胞器分布的细胞内极性得以维持。37℃下的悬浮细胞保持稳定的单价阳离子组成,但暴露于10(-5)M的哇巴因后,会迅速失去钾并获得钠。细胞内淀粉酶浓度以及细胞对肾上腺素或异丙肾上腺素的肾上腺素能刺激作出反应时淀粉酶的分泌模式和环磷酸腺苷的合成模式,与完整原位腺体的情况相当。此外,这些细胞表现出良好的氧气消耗,并在长达8小时的时间内保持恒定。这些细胞可作为实验工具,用于体外研究受体生理学和生物化学、细胞膜功能、细胞分泌机制以及外分泌腺细胞生理学的其他参数。