In Vitro Sustained Differentiation of Rat Colon Epithelial Stem Cells

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.

Design and Synthesis of Quinoxaline Hybrids as Modulators of HIF-1a, VEGF, and p21 for Halting Colorectal Cancer

“…All of the prepared quinoxaline derivatives were tested for their anticancer potential on colorectal cancer cells (HCT-116) via MTT assay. − Generally, most of them showed promising anticancer activity (Table ). Appealingly, compounds 6 , 9 , 14 , and 20 exhibited submicromolar IC 50 0.05–0.07 μM against HCT-116 cells within their safe dose (EC100), while DOX displayed IC 50 0.761 μM.…”

“…All of the compounds were safer than the reference doxorubicin (DOX, SI 0.357). Concerning EC100, all tested compounds, excluding 5 , 11 , 13 , and 21 showed a high safety profile on the growth of normal colonocytes with EC100 ≥ 0.1 μM − (Table ).…”

“…The normal colonic epithelial cells were isolated utilizing the Follmann & Birkner method 22 with modifications 23 and the cytotoxicity evaluation of the novel compounds were tested via MTT assay 24 as mentioned in the Supporting Information Data .…”