Schiewe M C, Schmidt P M, Bush M, Wildt D E
J Anim Sci. 1985 Jun;60(6):1610-8. doi: 10.2527/jas1985.6061610x.
Four hundred eight-cell mouse embryos were cultured in vitro in either polystyrene (plastic) dishes (Exp. 1) or watch glasses (Exp. 2) to analyze the toxicity potential of absorbed-retained ethylene oxide (EtO). Culture dishes were gas-sterilized with Anprolene equipment and allowed to aerate (21 C) for varying durations. Post-sterilization EtO residues, as determined by weight measurement, were eluted from polystyrene dishes at an exponential rate. After 1 wk of aeration, .625 mg EtO was retained/g of polystyrene product. To determine the effect of EtO residue on in vitro culture, embryos were collected from superovulated donor mice (C57BL/6N), pooled in a modified Dulbecco's phosphate buffered saline (PBI) medium and then randomly allotted to dishes subjected to various aeration durations. Embryos were cultured in Whitten's medium +3 mg/ml bovine serum albumin (BSA) under a humidified 5% CO2 in air atmosphere at 37 C. Gross assessments of embryo development were made using standard morphology and quality grading systems and a fluorescein diacetate staining assay. In Exp. 1, embryo development was retarded at the 8- to 16-cell stage when less than or equal to 12 h aeration of polystyrene dishes was permitted. Aeration for 24 and 36 h resulted in suboptimal embryo development with fewer (P less than .01) blastocysts and greater degeneration rates at 48 h of in vitro culture compared with the control (manufacturer-packaged dishes, no EtO) and 1-wk aeration treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)
400个八细胞期小鼠胚胎在聚苯乙烯（塑料）培养皿（实验1）或表面皿（实验2）中进行体外培养，以分析吸收留存的环氧乙烷（EtO）的潜在毒性。培养皿用安普罗林设备进行气体灭菌，并在不同时长下进行通气（21℃）。灭菌后通过重量测量确定的环氧乙烷残留量，以指数速率从聚苯乙烯培养皿中洗脱。通气1周后，每克聚苯乙烯产品留存0.625毫克环氧乙烷。为了确定环氧乙烷残留对体外培养的影响，从超排供体小鼠（C57BL/6N）收集胚胎，汇集于改良的杜氏磷酸盐缓冲盐水（PBI）培养基中，然后随机分配到经过不同通气时长处理的培养皿中。胚胎在惠顿培养基 + 3毫克/毫升牛血清白蛋白（BSA）中，于37℃、5%二氧化碳的湿润空气环境中培养。使用标准形态学和质量分级系统以及荧光素二乙酸酯染色试验对胚胎发育进行总体评估。在实验1中，当聚苯乙烯培养皿通气时间小于或等于12小时时，胚胎发育在8至16细胞阶段受到阻滞。与对照组（制造商包装的培养皿，无环氧乙烷）和通气1周的处理组相比，通气24小时和36小时导致胚胎发育欠佳，体外培养48小时时囊胚数量减少（P小于0.01），退化率更高。（摘要截断于250字）
