Abstract
Colonic epithelium composes of various cell types including alkaline phosphate-expressing absorptive, mucussecreting goblet and neuroendocrine cells that are derived from stem cells through asymmetric division. The continuous renewal of stem cells occurs under the highly coordinated cellular redox state. In the current study, based on a comparison with other culture media, colon epithelial cells were able to be sustained in vitro with normal status for more than two months under the chosen culture condition; α-MEM medium containing 20% fetal bovine serum. The cultured epithelial cells had normal doubling time and normal morphological characteristics as examined by transmission electron microscope. Also, these cultured cells contained functional stem cells and maintained their differentiation potency of colon stem cells, compared with freshly isolated mucosal epithelial cells, as indicated by the maintaining of aldehyde dehydrogenase 1B1 expression (11.31 ± 0.45 to 11.15 ± 0.48), ability to reduce silver nitrate, alkaline phosphate activity (0.513 ± 0.007 mU/μg to 0.438 ± 0.005 mU/μg), mucin secretion (34.71 ± 0.714 μg/ml to 32.93 ± 0.357 μg/ml) in appropriate cellular redox state level (-258.4 ± 1.3 mV to -237.4 ± 3.7 mV). The present study showed sustaining replication potential and functional differentiation of colonic epithelial stem cell population in this culture. The above culture system may be useful as an in vitro model for stemness, toxicological, and carcinogenesis studies.
Highlights
The glandular colonic epithelium composes of various cell types including absorptive, mucus-secreting goblet, and enteroendocrine or neuroendocrine cells
Doubling time of cells cultivated in α-MEM (3.14 ± 0.03) and Dulbecco's modified eagle's medium (DMEM) (5.23 ± 0.18) culture media showed a significant decrease compared to that of cells cultivated in Roswell Park Memorial Institute (RPMI) 1640 (8.47 ± 0.23) and Ham’s F-12/ DMEM (10.45 ± 0.27) culture media
These data were supported by healthy and ideal morphology of confluent epithelial cells cultivated in α-MEM and DMEM culture media as compared to that cultivated in RPMI and Ham’s F-12/ DMEM (1:1) culture media (Figure 2)
Summary
The glandular colonic epithelium composes of various cell types including absorptive (enterocytes), mucus-secreting goblet, and enteroendocrine or neuroendocrine cells. All these colonic epithelial cells derived from the stem cells [1]. Such stem cells can be defined by aldehyde dehydrogenase (ALDH) 1 that has been considered as a marker for normal human colonic stem cells. The continuous renewal of colonic stem cells is necessary for the maintenance of normal gut structure and function and it occurs through the highly coordinated and tightly regulated cellular redox state (Eh) that has been implicated in cell cycle responses such as proliferation, differentiation, and apoptosis. The dynamics of cellular redox balance are achieved by maintenance of the thiol (GSH)-todisulfide (GSSG) status [4]
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