Effects of Extracellular Matrix Proteins on Mouse Epididymal Epithelial Cell Attachment in Vitro

Objective: Appropriate extracellular matrix (ECM) molecules have been reported to produce significant improvement in the attachment, growth, and differentiation of many types of cell cultures. In this study, the effects of four ECM proteins (type IV collagen, type I collagen, fibronectin, and laminin) on the attachment of epididymal epithelial cells in culture were investigated. Materials and Methods: The epithelial cells were obtained from corpus epididymides of 4- and 6-week-old BALB/c mice and cultured with RPMI 1640 medium supplemented with 10% fetal calf serum in ECM-coated dishes at 37°C. The epithelial nature was determined by an indirect immunocytochemical staining technique. The attachment assay was performed and proteins in the cultured cells were analyzed by SDS-PAGE and autoradiography. Results: Type IV collagen was determined to significant improve the cell attachment whereas as a plastic surface without the ECM coating or coated with the remaining three ECM proteins exhibited relatively poor effects. Moreover, the epididymal epithelial cells from 4-week-old mice exhibited better attachment and proliferation than their 6-week-old counterparts with sperm in the epididymal tubules. On day 4, most of the cells revealed epithelial cell-specific keratins. WGA-binding proteins (GP-83 and GP-49) were found in the cell extracts and cultured media by isolation with WGA-agarose. However, there were no apparent differences in the pattern of protein synthesis between cells isolated from 4- and 6-week-old mice. Conclusions: This culturing model with epididymal cells recovered from the corpus epididymis of 4-week-old mice and cultured in type IV collagencoated dish is suitable for the investigation of epididymal physiology and sperm maturation in vitro.

並列關鍵字

epididymal cells ； fibronectin ； laminin ； type IV collagen

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