ISOLATION AND CULTURE OF EPITHELIAL-CELLS FROM RAT DUCTULI EFFERENTESAND INITIAL SEGMENT EPIDIDYMIDIS

To improve the study of epithelial function in rat ductuli efferentes (efferent ductules) and initial segment epididymis, we developed a pri mary cell culture system with modifications of the Klinefelter method (1992), The cultured efferent ductal epithelium was grown to confluenc e and the cells maintained many of the organelles characteristic of th ese cells in vivo, including dense-staining granules, indented nuclei and apical cilia, Ciliary beat was observed for up to 10 days in cultu re, Cultured initial segment epithelial cells were elongated and chara cterized by apical branched microvilli. Electron microscopy revealed i ntact cell junctions, an endocytotic apparatus and lysosomal granules, Ultrastructurally, the initial segment epithelium contained a well de veloped Golgi apparatus, For both epithelia, cell characteristics were also confirmed by indirect immunofluorescent staining for cytokeratin s 8, 18, Endocytotic activity was detected by the uptake of cationic f erritin at the apical surface and within vesicles, Estrogen receptor a nd clusterin mRNAs were expressed in the cultured epithelia and no dif ference was found in their expressions when cultured with or without 1 0(-9) M 17-beta estradiol, Indirect immunofluorescent staining for clu sterin further indicated that this protein was present in the cultures , In conclusion, these in vitro methods will be useful for the investi gation of epithelial function in the head of the epididymis.