Decline in Fluorescent Low Density Lipoprotein (LDL) Uptake by Small and Large Porcine Luteal Cells with Advancing Age of the Corpus Luteum1

Abstract

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)