IN-VITRO GROWTH OF CHLAMYDOPHILA ABORTUS IN OVINE ENDOMETRIUM: EVIDENCE OF GROWTH SUPPORT IN STROMAL FIBROBLAST CELLS
Keywords:Chlamydophila abortus, endometrium, epithelial cells, ovine, stromal fibroblast cells
The aim of present study was to develope an in vitro model to investigate the mechanism of Chlamydia abortus (C. abortus) infection in endometrial cells. The cells were isolated from uterine tissue of cyclic ewes (n=6) through digestion by collagenase alone or collegians plus trypsin. Digestion with collagenase alone yielded a lower epithelial-stromal cell ratio compared to digestion with collagenase and trypsin. Monolayers grown from cells using both enzymatic digestion methods were provisionally identified as epithelial or stromal fibroblast. The epithelial and fibroblast cell monolayers were infected with 50 µl of McCoy cell lysate containing 3x104 IFU/ml of C. abortus. No inclusions were observed in endometrial epithelial cells after 48-72 h post infection, however, typical inclusion bodies were observed in stromal fibroblasts. In vitro, C. abortus grew readily in endometrial stromal fibroblasts, the epithelial cells failed to support the growth of this organism. In conclusion, the disseverment protocols provide pristine populations of ovine endometrial epithelial and stromal cells and the cultured epithelial cells which may exhibit characteristics of in- vivo morphology and polarized function. This in-vitro method could accommodate as a consequential implement for further studies to investigate the different steroidal receptors in these cells at different stages of estrous cycle and their possible effects on susceptibility of the cells to C. abortus.
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