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A NOVEL SYSTEM FOR THE CO-CULTURE OF EPIDIDYMAL EPITHELIAL CELLS AND SPERM FROM ADULT RATS
Citation:
Klinefelter, G. A NOVEL SYSTEM FOR THE CO-CULTURE OF EPIDIDYMAL EPITHELIAL CELLS AND SPERM FROM ADULT RATS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-93/019 (NTIS PB93156248).
Description:
To study interactions which occur between the epididymal epithelial cells and sperm within the epididymis during sperm maturation, a specialized co-culture system capable of supporting the differentiated function of these cell types must be utilized. A multifaceted approach has been derived for isolation of plaques of epithelial cells from the epididymis of adult rats which involves microdissection, two collagenase digestions, and extensive extracellular matrix (ECM) overlying the filter within a primary semipermeable support forming into highly contiguous, but not confluent, regions of polarized epididymal epithelial cells during the first two days of culture. On the third day, immature sperm from the caput epididymidis are recovered and placed within a second semipermeable support resting above the epithelial cells. uring 24 hours of co-culture both sperm and epithelial integrity is maintained. oreover, certain functions requisite to epididymal sperm maturation occur such as protein secretion by the epithelial cells, the association of secreted protein with the sperm, and the acquisition of progressive motility by the sperm. omprehensive and illustrative description of this novel co-culture methodology is provided.