Selection of a chemically defined medium for culturing fetal mouse small intestine |
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Authors: | R Calvert P A Micheletti |
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Institution: | (1) Intestinal Biology Unit, Department of Anatomy and Cell Biology, Faculty of Medicine, University of Sherbrooke, J1H 5N4 Sherbrooke, Quebec, Canada |
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Abstract: | Summary We evaluated six commercially available tissue culture media in their capacity to support villi morphogenesis and enterocyte
differentiation during duodenal development of the fetal mouse in vitro: McCoy's 5A, Medium 199, Swim's S77, Trowell T8, Leibovitz
L-15, and RPMI-1640. The duodenal segments were resected at 15 d gestation, before the formation of intestinal villi. In the
segments cultured with the first four media, no villi differentiated even at 72 h culture. The number of epithelial cells
per transverse section of the explants did not increase at 24 h and thereafter the number of epithelial cells decreased, except
with McCoy's 5A. With the Leibovitz and RPMI media, rudimentary villi differentiated at 24 h of culture and they attained
their longest length at 48 h. With the RPMI medium, the number of epithelial cells doubled at 24 h of culture and with Leibovitz
medium it doubled at 48 h. At the fine structural level absorptive cells remained poorly differentiated with all the media
studied. Goblet cells were easily identified after 24 h culture; they had a well developed rough endoplasmic reticulum and
numerous mucous granules. Endocrine cells differentiated in culture and they were loaded with secretion granules. It was concluded
that the small intestine of the fetal mouse can be kept in organ culture for at least 72 h. Full maturation of absorptive
cells seemed to require some additional factor(s) as they remained poorly differentiated with all the media studied. Because
well differentiated endocrine cells were present in all the explants, it appeared that gastrointestinal hormones do not affect
villi morphogenesis and absorptive cells differentiation.
This investigation was supported by Grant MA-6069 from the Medical Research Council of Canada. Mr. P. A. Micheletti was supported
by a studentship from the FCAC. |
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Keywords: | organ culture fetal mouse intestine morphology cytochemistry |
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