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Title: GROWTH OF MANDUCA SEXTA MIDGUT EPITHELIAL CELLS IN THE ESTABLISHMENT OF A PRIMARY CULTURE

Author
item HAKIM, R - HOWARD UNIVERSITY
item HAKIM, F - NATIONAL INST OF HEALTH
item LOEB, MARCIA

Submitted to: Society for Invertebrate Pathology Meeting
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/12/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary: As an insect feeds, all of the nutritive material as well astoxins, bacteria and viruses are delivered to the midgut; nutrients aretaken into the midgut cells and thence transferred to the rest of the body. Similar paths are taken by the pathogens. Therefore, the gut is a most essential organ for maintaining the health of an insect, and also for the entrance of pathogenic organs. We have established a means of culturing normal midgut cells outside of the body. We have attempted to study these living cells in order to find clues to how the cells function in the whole animal. We report here that the cells change their nuclear DNA content with age, even though mature cells do not divide. This may imply that their function changes as they age inside the gut. This information will be useful to scientists learning how better to infect insects with pathogenic organisms which enter via the gut.

Technical Abstract: Primary cultures of Manduca sexta midgut epithelia are routinely prepared from cells collected from the pharate fourth instar midgut. These include rapidly proliferating populations of morphologically undifferentiated granular and smooth appearing spherical cells. Granularity is established as a cell feature in the flow cytometer. In established cultures over three weeks old we find mature differentiated cells - goblet and columnar cells- form along with stem cells, developing cells and dying cells. Cell cycling is reduced and the smooth cells contain primarily 2N DNA while the granular cells show a reduction in the 2N population, an increase in the 4N population, and small populations of cells of higher ploidy.