Figure 2.

Growth characteristics of WT-, EV-, and AS-ARH cells in vitro. (a) WT-, EV-, and AS-ARH cells (10⁵) were cultured in six-well plates containing RPMI-1640 media containing 10% FBS. At days 3 and 5 of the culture, the cells were sampled, stained with trypan blue, and counted. (b) ST2 cells (10⁴) in αMEM containing 10% FBS were plated onto dentin slices in 48-well plates. After 24 hours, WT-, EV-, and AS-ARH cells (10⁴) in RPMI-1640 media containing 10% FBS were added to the culture. At days 4 and 7 of the culture, viable cells were scored as described above. Growth rates of WT-, EV-, and AS-ARH cells were not significantly different in the presence or absence of ST2 cells cocultured on dentin slices. (c) Conditioned media from WT-, EV-, or AS-ARH cells (10⁵/ml) cultured in RPMI-1640 media containing 10% FBS were harvested at day 3, and the expression levels of MIP-1α were measured with a MIP-1α ELISA kit according to the manufacturer’s protocol. Similar results were seen in three independent experiments (*P < 0.0001).