Three-dimensional environments preserve extracellular matrix compartments of ovarian follicles and increase FSH-dependent growth.
In this study we performed a systematic comparative analysis of two culture environments-flat/adhesive liquid and three-dimensional collagen gel-upon in vitro ovarian follicle development. We paid particular attention to the effects of in vitro environments upon the preservation of follicular structure and of peri- and intra-follicular extracellular matrix. We show that flat/adhesive environment leads to an obvious distortion of follicle morphology, marked extracellular matrix modifications and high rates of spontaneous, i.e., FSH-independent, follicle disruption. In contrast, three-dimensional collagen gel environments are able to maintain follicular structure with an in vivo-like basal lamina architecture, minimizing spontaneous disruption. Follicle distortions found in flat/adhesive culture systems include a pronounced flattening, causing the follicle horizontal diameters not to adequately reflect follicle volume. Our volume data, based on three-axis follicle diameter measurements, indicate that three-dimensional collagen gel environments increase follicle growth, particularly in response to FSH. This study demonstrates that preservation of both peri- and intra-follicular extracellular matrix compartments during the in vitro growth and differentiation of ovarian follicles is highly desirable, and is now possible through the use of appropriate three-dimensional collagen gel culture environments. This system allows a better understanding of the specific roles played by each of the follicle compartments during development.
Gomes
J E
JE
Instituto de Histologia e Embriologia, Faculdade de Medicina de Lisboa, Lisboa Codex, Portugal.
Correia
S C
SC
Gouveia-Oliveira
A
A
Cidadão
A J
AJ
Plancha
C E
CE
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism* / Q000494:pharmacology; D006062:Gonadotropins / Q000502:physiology; D006107:Granulosa Cells / Q000502:physiology; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D008858:Microscopy, Phase-Contrast; D006080:Ovarian Follicle / Q000166:cytology* / Q000254:growth & development*
D006062:Gonadotropins; D005640:Follicle Stimulating Hormone; D003094:Collagen