Calvo L, Dennison-Lagos L, Banks S M, Fugger E F, Sherins R J
Division of Andrology, Genetics & IVF Institute, Fairfax, VA 22031.
Hum Reprod. 1993 Apr;8(4):575-80. doi: 10.1093/oxfordjournals.humrep.a138099.
We studied the effect of media composition on sperm capacitation, using Biggers-Whitten-Whittingham (BWW) medium, Ham's-F10 and a modified Tyrode's medium (HSM) supplemented with bovine serum albumin (BSA) or fetal cord serum (FCS). We evaluated the effect of chemical environment and protein supplementation on the sperm motion parameters of curvilinear velocity and linearity, and on the ability of incubated spermatozoa to undergo follicular fluid induced acrosome reaction. Neither chemical composition nor protein supplementation of capacitation media greatly affected motion parameters after 2 h incubation. Furthermore, chemical composition had only a small effect on the ability of spermatozoa to undergo the acrosome reaction upon exposure to follicular fluid. A higher proportion of spermatozoa underwent acrosome reaction after incubation in HSM (8% control (C); 28% follicular fluid) than in BWW (8% C, 17% follicular fluid) or Ham's F-10 (6% C, 19% follicular fluid). By contrast, protein source proved critical in determining acrosome reaction inducibility. Spermatozoa incubated in BSA-supplemented media showed a 4-fold increase in acrosomal discharge when exposed to follicular fluid (6% C, 22% follicular fluid) compared to controls while spermatozoa incubated in FCS were unable to undergo acrosome reaction (6% C, 6% follicular fluid). Simultaneous addition of FCS to BSA capacitation medium blocked acrosome reaction inducibility and the late addition of BSA, after sperm incubation in FCS, did not facilitate acrosome reaction. We propose that an inhibitor of sperm capacitation is present in FCS and therefore, the selection of optimum incubation conditions for spermatozoa may be of critical importance when evaluating or treating infertile patients.
我们使用比格斯 - 惠滕 - 惠廷厄姆(BWW)培养基、哈姆氏F - 10培养基以及添加了牛血清白蛋白(BSA)或胎牛血清(FCS)的改良泰勒氏培养基(HSM),研究了培养基成分对精子获能的影响。我们评估了化学环境和蛋白质补充对精子曲线速度和直线性等运动参数的影响,以及对孵育后的精子在卵泡液诱导下发生顶体反应能力的影响。在孵育2小时后,获能培养基的化学成分和蛋白质补充对运动参数的影响都不大。此外,化学成分对精子在接触卵泡液时发生顶体反应的能力影响也很小。与在BWW培养基(8%对照(C);17%卵泡液)或哈姆氏F - 10培养基(6% C,19%卵泡液)中孵育相比,在HSM培养基中孵育后有更高比例的精子发生顶体反应(8%对照(C);28%卵泡液)。相比之下,蛋白质来源在决定顶体反应诱导能力方面被证明至关重要。与对照组相比,在添加BSA的培养基中孵育的精子在接触卵泡液时顶体释放增加了4倍(6% C,22%卵泡液),而在FCS中孵育的精子则无法发生顶体反应(6% C,6%卵泡液)。向BSA获能培养基中同时添加FCS会阻断顶体反应诱导能力,并且在精子在FCS中孵育后再添加BSA也不会促进顶体反应。我们提出FCS中存在精子获能抑制剂,因此,在评估或治疗不育患者时,为精子选择最佳孵育条件可能至关重要。