Racowsky C, Kaufman M L, Dermer R A, Homa S T, Gunnala S
Department of Obstetrics and Gynecology, University of Arizona, Tucson 85724.
Fertil Steril. 1992 May;57(5):1026-33. doi: 10.1016/s0015-0282(16)55021-3.
To modify Tarkowski's air-dry technique for mouse oocytes to develop a rapid, consistent procedure for human oocytes that enables accurate scoring of meiotic stage.
DESIGN, SETTING, AND PATIENTS: Meiotically immature human oocytes, obtained after oophorectomy, were cultured for various periods and then subjected to Tarkowski's air-dry procedure (n = 104) or to our modified procedure (n = 175) that used a brief exposure to protease (20 to 40 seconds) before fixation.
Air-dried oocytes were assessed for readability and for whether they contained overspread or overlapping chromosomes. In addition, discrete meiotic stages in human oocytes were identified.
Our protease procedure significantly increased readability of air-dried oocytes (96% versus 79% readable for protease versus Tarkowski, respectively; P less than 0.001) by significantly reducing the number of preparations with either overscattered (0.7% versus 3.4% for protease versus Tarkowski, respectively, P less than 0.05) or overlapping (1.3% versus 18% for protease versus Tarkowski, respectively, P less than 0.001) chromosomes.
Protease exposure of oocytes, combined with a modification of Tarkowski's procedure, resulted in high quality air-dries of human oocytes. This rapid and reliable procedure should have clinical application in in vitro fertilization programs for meiotic assessment of oocytes failing to fertilize.
改进用于小鼠卵母细胞的塔科夫斯基空气干燥技术，以开发一种适用于人类卵母细胞的快速、一致的程序，从而能够准确划分减数分裂阶段。
设计、场所和患者：对卵巢切除术后获得的减数分裂未成熟人类卵母细胞进行不同时间段的培养，然后对其采用塔科夫斯基空气干燥程序（n = 104）或我们改进的程序（n = 175），后者在固定前短暂暴露于蛋白酶（20至40秒）。
评估空气干燥卵母细胞的可读性以及是否含有过度分散或重叠的染色体。此外，识别出人类卵母细胞中的离散减数分裂阶段。
我们的蛋白酶程序通过显著减少过度分散（蛋白酶组与塔科夫斯基组分别为0.7%对3.4%，P < 0.05）或重叠（蛋白酶组与塔科夫斯基组分别为1.3%对18%，P < 0.001）染色体的制备数量，显著提高了空气干燥卵母细胞的可读性（蛋白酶组与塔科夫斯基组可读率分别为96%对79%；P < 0.001）。
卵母细胞暴露于蛋白酶并结合对塔科夫斯基程序的改进，可实现高质量的人类卵母细胞空气干燥。这种快速且可靠的程序应在体外受精项目中具有临床应用价值，用于对未受精的卵母细胞进行减数分裂评估。
