Isolation, culture, and immunocytochemical characterization of epididymal epithelial cells from pubertal and adult rats.

Abstract

A method for the isolation and culture of epididymal e ithelial cells obtained from pubertal and old adult rats is d red. This method permits the establishment of primary cultures of these cells in monolayers from aggregates isolated from whole epididymides and major epididymal anatomical segments (caput, corpus, and cauda) after trypsin and collagenase digestions. A large number of cultured epididymal cells retain a differentiated function as demonstrated by the immunocytochemical and radioimmunoassay finding of acidic epididymal glycoprotein, a spermatozoa-coating protein secreted by the principal cells of rat epididymis. The proliferative tential of cultured epididymal cells obtained from pubertal an old adult donors can be documented by [3H]thymidine labeling and mitotic indices without significant loss of gene expression for acidic epididymal glycoprotein.Results of this study demonstrate that epididymal epithelial cells, consisting of a predominant population of principal cells, can be isolated, cultured, and maintained for up to 3 months.

Carnitine/organic cation transporter OCTN2-mediated transport of carnitine in primary-cultured epididymal epithelial cells

“…Epididymal epithelial cells were isolated from 5-week-old Sprague-Dawley rats (Saitama Experimental Animal Supply Co. Ltd, Saitama, Japan) according to the method reported previously (Kierszenbaum et al 1981, Leung et al 2001. To minimize contamination of non-epithelial cells and inhibition of attachment of epithelial cells to culture dishes by spermatozoa, we used immature rats, which do not contain spermatozoa (Kierszenbaum et al 1981, Leung et al 2001.…”

“…To minimize contamination of non-epithelial cells and inhibition of attachment of epithelial cells to culture dishes by spermatozoa, we used immature rats, which do not contain spermatozoa (Kierszenbaum et al 1981, Leung et al 2001. Briefly, epididymides were dissected out and minced into small fragments.…”

“…The resultant cell suspension was filtered through four sheets of gauze. Isolated epididymal cells were plated in tissue culture dishes at 5 £ 10 5 cells/ml and incubated at 32.5 8C for 10 h. Contaminating fibroblasts and smooth muscle cells became attached to the dishes within 10 h, so that epididymal epithelial cells could be separated from nonepithelial cells (Kierszenbaum et al 1981). The supernatant containing epididymal epithelial cells was collected and transferred to new dishes at 1 £ 10 5 cells/cm 2 .…”

In-vitro culture of hamster epididymal epithelium and induction of sperm motility

“…Since the cellular organization of an epithelium plays an integral part in its function, an important consideration in our culturing of the epididymal epithelium was to avoid procedures which initially reduce the tissue to a single cell suspension, as have been described elsewhere (Kierszenbaum et al, 1981;Klinefelter et al, 1982;Joshi, 1985). It may be argued that culture of intact lengths of epididymal tubules (Orgebin-Crist & Jahad, 1979) could prevent spermatozoa being exposed to a potentially activating environment.…”

“…Previous attempts to culture epididymal principal cells in vitro have shown that normal morphological features and some biochemical function (e.g. steroid synthesis) can be maintained for several days (Kierszenbaum et al, 1981;Klinefelter et al, 1982;Joshi, 1985). Furthermore, the organ culture of epididymal tubules, in which morphological integrity of epithelium remains largely intact, will induce some degree of sperm maturation (Orgebin-Crist & Jahad, 1979;Cuasnicu et al, 1984), but this culture methods allows only limited access to the tubule lumen.…”