Characterization of kidney epithelial cells from the Florida manatee, Trichechus manatus latirostris |
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Authors: | Sweat JMDunigan D D Wright S D |
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Institution: | (1) Marine Mammal Pathobiology Laboratory, Florida Fish and Wildlife Conservation Commission, 3700 54th Avenue South, 33711 St. Petersburg, Florida;(2) Department of Pathobiology, College of Veterinary Medicine, University of Florida, 2015 SW 16th Avenue, 32610 Florida, Gainesville;(3) Gulf Coast Research and Education Center, University of Florida, 13138 Lewis-Gallager Road, 33527 Dover, Florida |
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Abstract: | Summary The West-Indian manatee,Trichechus manatus latirostris, is a herbivorous marine mammal found in the coastal waters of Florida. Because of their endangered status, animal experimentation
is not allowed. Therefore, a cell line was developed and characterized from tissue collected during necropsies of the manatees.
A primary cell culture was established by isolating single cells from kidney tissue using both enzymatic and mechanical techniques.
Primary manatee kidney (MK) cells were subcultured for characterization. These cells were morphologically similar to the cell
lines of epithelial origin. An immunocytochemistry assay was used to localize the cytokeratin filaments common to cells of
epithelial origin. At second passage, epithelial-like cells had an average population-doubling time of 48 h, had an optimum
seeding density of 5×103 cells/cm2, and readily attached to plastic culture plates with a high level of seeding efficiency. Although the epithelial-like cells
had a rapid growth rate during the first three passages, the cloning potential was low. These cells did not form colonies
in agar medium, were serum dependent, had a limited life span of approximately nine passages, and possessed cell-contact inhibition.
These data suggest that the cells were finite (noncontinuous growth), did not possess transformed properties, and were of
epithelial origin. These cells are now referred to as MK epithelial cells. |
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Keywords: | Florida manatee cell culture cytokeratins endangered species |
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