Todd M E, Friedman S M
In Vitro. 1978 Sep;14(9):757-70. doi: 10.1007/BF02617969.
The rat-tail artery was maintained in vitro for 2 weeks to investigate its suitability as an experimental model. The criteria were that (a) it should retain the overall histological organization with normal ultrastructural appearance of the smooth-muscle cells; (b) stored neurotransmitter which could be activated by experimental treatment should be absent; and (c) smooth-muscle ion transport mechanisms should fall within normal range. Vessels were maintained in Falcon tissue-culture dishes in Dulbecco's modified Eagle's medium. Either 2% or no serum supplement was found to be more suitable than 10% serum due to the high rate of cell proliferation induced by the latter. Light and electron microscopy of cross sections of the vessels indicated that the overall normal vessel architecture was retained, and the ultrastructural features predicted normal function. There were no discernible differences dependent on the length (up to 8- to 10-cm lengths) of the cultured vessel. Preliminary experiments with fluorescent microscopy showed that stored neurotransmitter in the nerves of the vessel wall was no longer present after 48 hr. Ultrastructural examination revealed that storage vesicles in vitro lost their dense cores, representing noradrenalin, between 41 and 48 hr in culture. Normal ion transport mechanisms were retained in the smooth-muscle cells of the arteries in vitro for up to 2 weeks when tested with ion-specific electrodes. Morphological and physiological evidence support the suitability of the rat-tail artery as a model for experimental testing of vascular tissues.
将大鼠尾动脉在体外维持2周,以研究其作为实验模型的适用性。标准如下:(a) 它应保留整体组织学结构,平滑肌细胞具有正常的超微结构外观;(b) 不存在可被实验处理激活的储存神经递质;(c) 平滑肌离子转运机制应在正常范围内。血管在Falcon组织培养皿中,置于杜氏改良伊格尔培养基中培养。由于10%血清诱导的细胞增殖率高,发现2%血清或无血清补充比10%血清更适合。对血管横截面的光镜和电镜检查表明,整体正常的血管结构得以保留,超微结构特征预示功能正常。培养的血管长度(最长8至10厘米)不同,未见明显差异。荧光显微镜初步实验表明,血管壁神经中储存的神经递质在48小时后不再存在。超微结构检查显示,培养41至48小时后,体外储存囊泡失去了代表去甲肾上腺素的致密核心。当用离子特异性电极测试时,体外动脉平滑肌细胞在长达2周的时间内保留了正常的离子转运机制。形态学和生理学证据支持大鼠尾动脉作为血管组织实验测试模型的适用性。