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題 名 | Effects of Extracellular Matrix Proteins on Mouse Epididymal Epithelial Cell Attachment in Vitro=細胞外基質蛋白對體外培養小鼠副睪上皮細胞附著的影響 |
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作 者 | 孫光煥; 劉鴻文; 吳勝堂; 于大雄; 閻中原; 陳宏一; 張聖原; | 書刊名 | Journal of Medical Sciences |
卷 期 | 22:5 2002.10[民91.10] |
頁 次 | 頁199-206 |
分類號 | 361.8 |
關鍵詞 | 基質蛋白; 副睪; 上皮細胞; Epididymal cells; Fibronectin; Laminin; Type Ⅳ collagen; |
語 文 | 英文(English) |
英文摘要 | 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 culture were investigated. Materials and Methods: The epithelia 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 disphes 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 th 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-wek-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 collagen-coated dish is suitable for the investigation of epididymal physiology and sperm maturation in vitro. |
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