Partial analysis of sequences expressed by human osteoblasts in vitro using differential display-PCR (DD-PCR)
Bone mineralization is a complex process that has not been fully elucidated. Human osteoblasts isolated from hip explants were cultured with (OB+) and without (OB) beta-glycerol phosphate ($\beta$GP), a promoter of mineralization. Complementary DNA from OB and OB+ cells was amplified by the differential display polymerase chain reaction (DD-PCR). This methodology identified a DD-PCR fragment from mineralized osteoblasts that was absent from non-mineralized cells. This fragment detected two different cDNAs in an OB+ cDNA library screen. One cDNA matched an EST and was expressed in many tissues, including both OB and OB+ RNA, although preliminary data indicated it may be expressed more highly in OB+ than OB cells. The second cDNA matched human mitochondrial DNA. The initial DD-PCR fragment was also cloned directly and yielded one potential recombinant of the correct size. However, the sequence encoded by this clone was not detected in osteoblasts by northern blot analysis.
Hill, Jason Eric, "Partial analysis of sequences expressed by human osteoblasts in vitro using differential display-PCR (DD-PCR)" (1996). ProQuest ETD Collection - Thomas Jefferson University. AAI1381336.