Primary EO cells were isolated from the lower incisors of Sprague-Dawley rats (100 to 120 g). Secretory and maturation EO cells were isolated as previously described using a molar reference line (38, 39). EO cells were digested with Liberase (0.25 mg/ml; Roche) for 30 min at 37°C, washed in Hanks’ balanced salt solution, and plated onto CellTak-coated (Corning) coverslips in X-Vivo15 medium (Lonza) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 1% glutamine. Isolated EO cells were used within 24 hours after dissection. LS8 cells are an immortalized murine-derived enamel cell line (37). LS8 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS at 37°C with 5% CO2.

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