男性不妊に関する研究 --in vitroにおけるヒト精子acrosome reaction--

Abstract

triple-stain techniqueを用いヒト精子acrosome reactionを観察した.1) mBWW中では培養6時間後まではacrosome reaction陽性精子の割合は漸増した.2) human serum albuminを含まない培養液中ではacrosome reactionはおこらなかった.Caイオン, Ca ionophore A23187, trypsinにはacrosome reactionの誘発作用を認めた.3)不妊患者精子の中で乏精子症患者の精子や運動率が20%以下の精子はFertile Maleの精子に比しacrosome reactionをおこさせると困難であった.これが不妊症の一因となっている可能性を示した

In 1981, P. Talbot developed a triple-stain technique to estimate the number of human sperm undergoing normal acrosome reaction in fixed smears. In this method, live and dead sperm are first differentiated using the vital stain trypan blue. Sperm are then fixed in glutaraldehyde, dried onto slides, and the postacrosomal region and acrosome are differentiated using Bismark brown and Rose Bengal. Slides are examined at 1, 000x with a bright-field microscope and assessed for the percentage of sperm that have undergone the normal acrosome reaction. Using this method we examined the time course of the acrosome reaction of human sperm incubated in mBWW the influence of human serum albumin, the calcium concentration of incubation media, Ca ionophore A 23187, and trypsin on the acrosome reaction of human sperm and the difference between the percentage of sperm undergoing normal acrosome reaction of fertile and infertile males. We got the following results: The percentage of human sperm undergoing acrosome reaction increased for the first six hours of incubation. Without human serum albumin the acrosome reaction did not occur. Ca ion could be one of the triggers of the acrosome reaction. Ca ionophore A23187 and trypsin induced the acrosome reaction in vitro. Sperm from oligozoospermic males, especially poorly motile sperm, could not undergo acrosome reaction so easily as sperm from fertile males could.