Adickes E D, Mollner T J, Lockwood S K
Research Service (151), Omaha Veterans Administration Hospital, NE 68105.
Alcohol Alcohol. 1988;23(5):377-81. doi: 10.1093/oxfordjournals.alcalc.a044832.
The accuracy and consistency of the delivery of ethanol to cultured cells is important to determine effects on morphologic, biochemical and physiologic alterations. Open and closed chamber systems were evaluated to determine cytotoxic vs sublethal, potentially teratogenic effects on neonatal rat cardiac myocytes. The open system employed a variety of cell culture vessels. Cardiac cells were exposed directly to ethanol in the growth media at concentrations of 5-50 mM in Petri dishes, multiwell slides and multiwell chambers. Ethanol concentrations in the media in these open vessels decreased over 60% in a 24 hr incubation period. A closed system consisted of tightly sealed plastic containers in which the same vessels were used. The vessels were placed on a platform over a bath of ethanol-water. Cells were acclimated for 24 hr with ethanol in the bath at 200% of the final desired media concentration. Ethanol gradually diffused into the media to reach peak levels of 5, 10, 25 or 50 mM at 24 hr. After the 24 hr period, ethanol was added to both the media and bath at the desired concentration. Cells exposed gradually to ethanol in the closed chambers remained viable, but showed slower division and growth. A period of gradual acclimation is required to induce sublethal cellular effects rather than lethal effects. The diversity of cell systems and manipulations of cultures to study the potential teratogenic effects of ethanol are improved using such a closed chamber system.
将乙醇递送至培养细胞的准确性和一致性对于确定其对形态学、生物化学和生理学改变的影响非常重要。对开放和封闭腔室系统进行了评估，以确定对新生大鼠心肌细胞的细胞毒性与亚致死性、潜在致畸作用。开放系统使用了多种细胞培养容器。在培养皿、多孔载玻片和多孔培养室中，心脏细胞直接暴露于生长培养基中浓度为5 - 50 mM的乙醇中。在24小时的孵育期内，这些开放容器中培养基中的乙醇浓度下降超过60%。封闭系统由紧密密封的塑料容器组成，其中使用相同的容器。容器放置在乙醇 - 水浴上方的平台上。细胞先用最终所需培养基浓度200%的乙醇在水浴中驯化24小时。乙醇逐渐扩散到培养基中，在24小时时达到5、10、25或50 mM的峰值水平。24小时后，以所需浓度将乙醇添加到培养基和水浴中。在封闭腔室中逐渐暴露于乙醇的细胞保持存活，但显示出较慢的分裂和生长。需要一段逐渐驯化的时间来诱导亚致死性细胞效应而非致死效应。使用这种封闭腔室系统可改善用于研究乙醇潜在致畸作用的细胞系统多样性和培养操作。
