Session: MON 515-547-Female Reproductive Endocrinology
Poster Board MON-528
DESIGN: Prospective study
MATERIAL AND METHODS: CCs and MGCs from 16 lean IVF women were studied. Cells were pooled by cell type, with each type of cell separated into two groups for determination of initial lipid content (Method [M]1) and lipid accumulation in vitro (Method [M]2). Cells for initial lipid content were immediately fixed at oocyte retrieval with 4% paraformaldehyde in suspension; those for lipid accumulation in vitro were cultured for 4 hours with 5% fetal calf serum and fixed. Cells were treated with lipid fluorescent dye BODIPY® FL C16and nuclear marker DAPI. Cell lipid was quantified by confocal microscopy with ImageJ software. Two-way ANOVA compared cell lipid and area by cell type and preparation; paired T-test compared CC/MGC lipid ratio between cell preparations; linear regression correlated cell lipid and area with patient BMI, age and gonadotropin dose.
RESULTS: There was no effect of cell type (P=0.2) or cell type-cell preparation interaction (P=0.8) on cell area (M1: CC 99.7 ± 5.1, MGC 132.8 ± 5.8; M2: CC 221.9 ± 30.4, MGC 265.1 ± 48.5 µm2). Mean area of all cells combined was significantly less for cells prepared by M1 (116.2 ± 4.9 µm2) vs. M2 (243.5 ± 22.5 µm2, P<0.00005). Cell lipid level, however, was altered by cell preparation (P<0.05; cell preparation-cell type interaction, P<0.00001). Initial lipid content was lower in CCs (74.5 ± 9.3) than MGCs (136.3 ± 16.7 fluorescence/cell area, P<0.00005), while lipid accumulation in vitro was higher in CCs (154.0 ± 9.1) than MGCs (104.6 ± 9.9 fluorescence/cell area, P<0.00001, cell type effect). The relatively diminished initial CC lipid content vs. CC lipid accumulation in vitro (P<0.00001), and the opposite pattern for MGCs (P<0.05 cell preparation effect), lowered the CC/MGC lipid ratio in M1 (0.55 ± 0.04) vs. M2 (1.58 ± 0.10, P<0.00001). Cell area and lipid amounts did not significantly correlate with patient BMI, age or amount of gonadotropin given.
SUMMARY: Ovarian cell lipid content of lean IVF women is spatially distributed at oocyte retrieval and altered by cell preparation, independent of patient characteristics within the ranges studied.
CONCLUSIONS: Differential uptake or utilization of CC and MGC lipid during human oocyte maturation and steroidogenesis is the function of both the follicular environment and the capacity of these cells to accumulate lipid in vitro. By characterizing normal ovarian cell lipid metabolism in lean IVF patients by different methods, future studies can explore the adverse effects of ovarian cell lipotoxicity on oocyte competence (3).
Disclosure: DHA: Ad Hoc Consultant, Viamet Pharmaceuticals, Inc.. Nothing to Disclose: PS, MA, EK, AS, PA, CB, DH, GDC, DAD
*Please take note of The Endocrine Society's News Embargo Policy at http://www.endo-society.org/endo2013/media.cfm
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