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S. Shimmura, H. Miyashita, T. Kawakita, N. Goda, M. Suematsu, K. Tsubota; Metabolomic Analysis of Calcium-Induced Differentiation in Murine Corneal Epithelial Cell Progenitors. Invest. Ophthalmol. Vis. Sci. 2007;48(13):446.
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To compare the metabolic profile in murine corneal epithelial progenitor cells (TKE2 clone) following calcium-induced differentiation.
Murine corneal epithelial progenitor cells (TKE2) were cultured and passaged in serum-free low calcium medium (defined KSFM, Invitrogen). TKE2 cells were induced to differentiate by adding 0.9 mM Ca2+ in the medium. Differentiation was confirmed by the expression of involucrin by RT-PCR. Metabolomic profiles of TKE2 and calcium-treated TKE2 cells were differentially displayed (metabolomic analysis) by coupling capillary electrophoresis with electrospray ionization time-of-flight mass spectrometry (CE-TOFMS).
The addition of calcium induced differentiation of TKE2 cells shown by the upregulation of involucrin. A total of 36 anions, 56 cations and 31nucleotides were measured by CE-TOFMS. The majority of metabolites analyzed increased following the addition of calcium in the medium. Metabolites increasing by more than two fold included pyruvate, lactate, fumarate, arginosuccinate, fructose 1,6-diphosphate, adenosine, guanosine, AMP, GMP and NADH.
Calcium-induced differentiation of TKE2 cells involved the increase of major metabolites of the oxydative phosphorylation and glycolysis pathway. A general shift was observed from nucleic acid synthesis to energy metabolism.
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