Micro-dissection efficiently separates different growth plate zones from cultured tibiae. E15.5 tibiae that were harvested and incubated with or without CNP (1 μM) for six days were micro-dissected into the resting/proliferating, hypertrophic, and mineralized regions as shown (A). Zones from approximately 24 bones were pooled together. RNA was isolated directly from micro-dissected tibia and analyzed by microarray as described in Materials and Methods. Real-time PCR analyses confirmed expected expression patterns of the cartilage markers Col2a1 and Col10a1 in control bones (B; data represent means ± SD from three independent trials). Expression patterns of selected chondrocyte marker genes under control conditions in our microarray data sets further demonstrated efficient separation of regions (C).
Agoston et al. BMC Developmental Biology 2007 7:18 doi:10.1186/1471-213X-7-18