Effects on gene expression of continuous exposure of mineralizing NHBC cultures to rhSCL.

NHBC were cultured under mineralizing conditions for up to 35 days in the absence or presence of rhSCL at 50 ng/ml. Media were replenished every 3–4 days. At the time points indicated, total RNA was prepared and real-time RT-PCR performed to determine mRNA expression of A) DMP1, B) SOST, C) OPG and D) RANKL. Data shown are means of triplicate reactions ± SD normalized to expression of 18S mRNA. Significant differences to untreated control are indicated by * p<0.05 and *** p<0.001. Similar results were obtained from 3 independent experiments using NHBC from different donors.