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Production of Gelatinases and Tissue Inhibitors of Matrix Metalloproteinases by Equine Ovarian Stromal Cells In Vitro¹

^a Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario,Canada N1G 2W1

Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) play very important roles in extracellular matrix (ECM) remodeling in ovarian follicle growth and ovulation. Equine follicles are embedded in cortex that is at the center of the ovary, and they must expand/emigrate to the fossa, the only site in the ovary for ovulation. Therefore, equine ovarian stromal cells (EOSC) are probably involved in ECM remodeling during follicle growth. This study examined whether cultured EOSC synthesize gelatinases and TIMPs, molecules essential for ECM remodeling in other systems. Results showed that cultured EOSC (passage 3–8) had a fibroblast-like morphology and were positive for -smooth muscle actin and type I procollagen by immunostaining. Gelatinase A (MMP-2), gelatinase B (MMP-9), TIMP-1, and TIMP-2 were present in EOSC-conditioned medium, and TIMP-3 in ECM of EOSC. Transforming growth factor ß significantly stimulated the activity of gelatinases A and B and TIMP-1 in conditioned medium from EOSC (p < 0.05). Phorbol 12-myristate 13-acetate also significantly stimulated the activity of gelatinases A and B and TIMP-1 in conditioned medium and of TIMP-3 in ECM (p < 0.05). Our results suggest that EOSC produce important components of the ECM remodeling machinery and, therefore, may play a role in the ECM remodeling during follicle growth in this species.

¹ Financial support provided by the Natural Sciences and Engineering Research Council of Canada (NSERC), the Dynasty Equine Trust Fund, and the Ontario Ministry of Agriculture, Food and Rural Affairs.

² Correspondence. FAX: 519 767 1450; bcoomber{at}uoguelph.ca

³ Deceased.

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