Table 2.

Evaluation of the histological staining results of D17 osteosarcoma cells grown as two‐dimensional (monolayer) and three‐dimensional (spheroid) cell culture

	Monolayer 2 day	Spheroid 7 day	Spheroid 14 day	Spheroid 19 day
Histochemical staining
Aniline blue	−/+	+	+	+
Periodic acid–Schiff	−/+	+	+	+
Safranin O	−	−	−	−
Immunohistochemical staining
Alkaline phosphatase	+	+	+	+
Ezrin	+	+	+	+
S100A4	+	+	+	+
Ki‐67	+	+	+	+
Collagen I	−/+	+	+	+
Cytokeratin	−	−	−	−
Vimentin	+	+	+	+
Osteocalcin	−/+	−/+	+	+

Scoring of the histochemical and immunohistochemical staining pattern: positive immunoreaction (+); no positive immunoreaction (−); (−/+) ranging from no positive to minor immunoreaction.