HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes

Figure 2

Proliferation and differentiation of HaCaT cells in low (0.07 mM) and high (1.8 mM) Ca²⁺-containing medium. (a) Proliferation of HaCaT cells plated at the same density (1.0 × 10⁴ cells/cm²) assessed by the MTT assay at 2, 6, 9, and 14 days of incubation. Values represent mean ± SD of three independent experiments. (b) Western blot analysis of the expression of keratinocyte (KC) differentiation markers (K10, K14, and involucrin) in HaCaT cells grown in low (a) and high (c) Ca²⁺-containing medium for 6 (A6 and C6) or 14 (A14 and C14) days. The relative intensities of band signals quantified by digital scanning densitometry are reported in the histogram; β-actin was used to normalize the results to protein content. This blot is a representative of three independent experiments. (c) Immunofluorescence staining of HaCaT cells, grown in low (A) and high (C) Ca²⁺-containing medium for 6 (A6 and C6) and 14 (A14 and C14) days, and of primary human KC, grown in low Ca²⁺-containing medium, with anti-K10 antibodies (magnification: 60x).

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