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人精子与人类输卵管上皮细胞单层共培养后的功能:研究人类早期受孕中细胞相互作用的体外模型

Human sperm function after coculture with human fallopian tubal epithelial cell monolayers: in vitro model for studying cell interactions in early human conception.

作者信息

Bongso A, Ho J, Fong C Y, Ng S C, Ratnam S

机构信息

Department of Obstetrics/Gynaecology, National University Hospital, Singapore.

出版信息

Arch Androl. 1993 Nov-Dec;31(3):183-90. doi: 10.3109/01485019308988398.

Abstract

It has been difficult to study the behavior of sperm in the tubal environment in vivo. Human tubal epithelial cells were therefore cultured in vitro to simulate tubal conditions and human sperm function was assessed in the presence of such cells in vitro. Ampullary epithelial cell lines were established from fallopian tubes collected from premenopausal women undergoing hysterectomy. Approximately 1 x 10(5) cells/mL from monolayers of the third passage were seeded with 1 mL of culture medium into each well of 4-well plastic dishes. Sperm from 10 ejaculates of 10 different oligoasthenozoospermic men 30-41 years of age were recovered by the swim-up method and 200,000 sperm from each ejaculate were added into each well at the time of cell seeding. Control wells were treated the same but without cells. All dishes were incubated at 37 degrees C in 5% CO2, and sperm motility, acrosome reaction, and sperm-cell binding assessed at 1, 5, and 24 h. Curvilinear velocity and mean amplitude of lateral head displacement were significantly different in ampullary cultures as compared with controls for all three time periods: 1 h (67 +/- 5.2 vs 58 +/- 4.9 microns/s, p < 0.05; 4.48 +/- 0.4 vs 3.29 +/- 0.3 microns; p < 0.05), 5 h (75 +/- 5.8 vs 64 +/- 5.0 microns/s, p < 0.05; 4.92 +/- 0.5 vs 3.68 +/- 0.3 microns, p < 0.05), and 24 h (70 +/- 4.8 vs 59 +/- 4.2 microns/s, p < 0.05; 4.36 +/- 0.4 vs 3.11 +/- 0.3 microns, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

在体内研究精子在输卵管环境中的行为一直很困难。因此,体外培养人输卵管上皮细胞以模拟输卵管条件,并在体外此类细胞存在的情况下评估人精子功能。从接受子宫切除术的绝经前妇女的输卵管中建立壶腹上皮细胞系。将第三代单层培养物中约1×10(5)个细胞/mL与1 mL培养基接种到4孔塑料培养皿的每个孔中。通过上游法收集10名年龄在30 - 41岁的不同少弱精子症男性的10次射精中的精子,并在接种细胞时将每次射精的200,000个精子添加到每个孔中。对照孔进行相同处理但不添加细胞。所有培养皿在37℃、5%二氧化碳条件下孵育,并在1、5和24小时时评估精子活力、顶体反应和精子 - 细胞结合情况。在所有三个时间段,与对照相比,壶腹培养物中的曲线速度和头部侧向位移平均幅度均有显著差异:1小时(67±5.2对58±4.9微米/秒,p<0.05;4.48±0.4对3.29±0.3微米;p<0.05),5小时(75±5.8对64±5.0微米/秒,p<0.05;4.92±0.5对3.68±0.3微米,p<0.05),以及24小时(70±4.8对59±4.2微米/秒,p<0.05;4.36±0.4对3.11±0.3微米,p<0.05)。(摘要截断于250字)

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