Dharmarajan A M, Yoshimura Y, Sueoka K, Atlas S J, Dubin N H, Ewing L L, Zirkin B R, Wallach E E
Johns Hopkins University, School of Medicine, Department of Gynecology and Obstetrics, Baltimore, Maryland 21205.
Biol Reprod. 1988 Jun;38(5):1137-43. doi: 10.1095/biolreprod38.5.1137.
An ovarian in vitro perfusion method was adapted to examine rabbit corpus luteum (CL) function during pseudopregnancy. Ovaries were perfused in vitro with tissue culture Medium 199 with or without 3% bovine serum albumin (BSA). Samples were obtained from both arterial and venous cannulae, allowing rates of progesterone secretion to be determined. Two perfusion methods were compared: a closed system in which perfusion medium continuously recirculated through the tissue, and an open system in which the ovarian vein cannula was left outside the perfusion chamber. The addition of 3% BSA was found to prevent edema and distortion of the interstitial space, and to result in increased progesterone secretion. With the closed perfusion system, the progesterone secretion rate measured over the 6-h perfusion was significantly higher on Day 11 than on Day 1 of pseudopregnancy and had declined significantly on Day 18. There was no difference in secretion rates whether the perfusion system was open or closed, and the results obtained with both systems closely approximated in vivo progesterone secretion rates. Measurements of CL tissue progesterone content before and after in vitro perfusion indicated that the changes in progesterone secretion seen during pseudopregnancy resulted from differences in the synthesis and secretion of progesterone and not from leakage of progesterone already present in the CL prior to perfusion. Taken together, these results indicate that the modified in vitro perfused rabbit ovary preparation described herein is an appropriate model to examine progesterone secretion by ovaries bearing CL.
采用一种卵巢体外灌注方法来检测假孕期间兔黄体（CL）的功能。将卵巢置于体外，用含有或不含有3%牛血清白蛋白（BSA）的199组织培养基进行灌注。从动脉插管和静脉插管中获取样本，从而测定孕酮分泌率。比较了两种灌注方法：一种是封闭系统，灌注培养基在组织中持续循环；另一种是开放系统，卵巢静脉插管留在灌注室外部。发现添加3% BSA可防止间质间隙水肿和变形，并导致孕酮分泌增加。在封闭灌注系统中，假孕第11天6小时灌注期间测得的孕酮分泌率显著高于假孕第1天，而在第18天则显著下降。无论灌注系统是开放还是封闭，分泌率均无差异，且两种系统获得的结果与体内孕酮分泌率非常接近。体外灌注前后CL组织孕酮含量的测量表明，假孕期间观察到的孕酮分泌变化是由孕酮合成和分泌的差异引起的，而非灌注前CL中已存在的孕酮泄漏所致。综上所述，这些结果表明本文所述的改良体外灌注兔卵巢制备方法是检测带有CL的卵巢孕酮分泌的合适模型。
