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Heui Uhm, Sheyla Gonzalez, Elfren Ray Baclagon, Sophie Xiaohui Deng; Inhibition of Notch via SAHM1 in human limbal epithelial stem/progenitor cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4908.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the effect of Notch signaling inhibition on human limbal epithelial stem/progenitor cells (LSCs) using a stapled α-helical peptide derived from MAML1 (SAHM1) in vitro.
Single primary human LSCs were isolated and cultured on 3T3 feeder cells in supplemented hormonal epithelial medium for up to 2 weeks. SAHM1, an inhibitor of Notch active transcriptional complex, was added to the culture at the concentration of 1, 5, 10 and 20 µM. Cultured LSCs were analyzed for cell morphology, cell expansion rate, colony forming efficiency (CFE), cell size, and expression levels of putative stem cell markers.
Morphology, cell size, and CFE of LSCs cultured in the presence of SAHM1 at all four concentrations were similar to those in the control. The cell expansion rate was reduced at high concentrations of SAHM1 (20 µM, p<0.05). The percentage of K14+ cells remained comparable for all conditions (p>0.05). The percentage of K12+ cells was lower at higher concentrations of SAHM1 (5, 10, and 20 µM) in comparison to the control (p<0.05). No significant differences were found in the percentage of cells expressing high levels of p63α in the presence of SAHM1.
Inhibition of Notch signaling using SAHM1 appears to reduce maturation of LSCs in vitro.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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