Brannian J D, Kurz S G, Shiigi S M
Department of Obstetrics and Gynecology, University of South Dakota School of Medicine, Sioux Falls 57117.
Biol Reprod. 1994 Jan;50(1):204-9. doi: 10.1095/biolreprod50.1.204.
The present study was designed to test the hypothesis that the ability of small luteal cells (SLC) and/or large luteal cells (LLC) to take up low density lipoprotein (LDL) declines with advancing age of the CL. Ovaries from 100-110-kg gilts were classified as early (Days 4-6; n = 5), mid (Days 8-12; n = 6)-, or late (Days 15-18; n = 5) cycle on the basis of gross morphology. Multiple CL from each ovary were pooled and enzymatically dissociated. An aliquot of dispersed luteal cells was reserved for cell culture. Remaining cells were incubated (approximately 4 x 10(5) cells/0.25 ml Dulbecco's Modified Eagle's Medium [DMEM] + 0.1% BSA) for 20 min at 37 degrees C with human LDL (10 micrograms/ml) tagged with the fluorescent probe, Dil (Dil-LDL). Washed and fixed cells were then isolated by flow cytometry into SLC and LLC subpopulations on the basis of forward and 90 degrees light scatter. Cellular fluorescence was analyzed within each subpopulation. The percentage of fluorescent, i.e., Dil-LDL-positive (+), SLC did not differ between early (29.8 +/- 5.9%) and mid (40.5 +/- 6.8%)-cycle, but declined (p < 0.01) in late CL (7.0 +/- 1.6%). Similarly, the percentage of Dil-LDL-(+) LLC was unchanged between early (80.5 +/- 2.0%) and mid (78.6 +/- 4.2%)-cycle, but diminished (p < 0.01) in late (40.2 +/- 1.9%) CL. Moreover, the percentage of total cells isolated in the LLC subpopulation declined dramatically (p < 0.01) between mid (8.0 +/- 0.9%)- and late (1.6 +/- 0.2%) cycle, but the percentage of SLC did not change.(ABSTRACT TRUNCATED AT 250 WORDS)
随着黄体（CL）年龄的增长，小黄体细胞（SLC）和/或大黄体细胞（LLC）摄取低密度脂蛋白（LDL）的能力会下降。根据大体形态，将100 - 110千克的后备母猪卵巢分为早期（第4 - 6天；n = 5）、中期（第8 - 12天；n = 6）或晚期（第15 - 18天；n = 5）周期。将每个卵巢的多个CL合并并进行酶解。取一份分散的黄体细胞用于细胞培养。其余细胞（约4×10⁵个细胞/0.25毫升杜尔贝科改良 Eagle 培养基[DMEM] + 0.1%牛血清白蛋白）在37℃下与人LDL（10微克/毫升）标记的荧光探针Dil（Dil - LDL）孵育20分钟。然后通过流式细胞术根据前向散射光和90度散射光将洗涤并固定的细胞分离为SLC和LLC亚群。分析每个亚群内的细胞荧光。荧光即Dil - LDL阳性（+）的SLC百分比在早期（29.8±5.9%）和中期（40.5±6.8%）周期之间无差异，但在晚期CL（7.0±1.6%）中下降（p < 0.01）。同样，Dil - LDL（+）的LLC百分比在早期（80.5±2.0%）和中期（78.6±4.2%）周期之间无变化，但在晚期（40.2±1.9%）CL中减少（p < 0.01）。此外，在LLC亚群中分离出的总细胞百分比在中期（8.0±0.9%）和晚期（1.6±0.2%）周期之间显著下降（p < 0.01），但SLC百分比没有变化。（摘要截断于250字）
