Three isolation techniques for primary culture of human osteoblast-like cells: A comparison

Osteolineage niche cells initiate hematopoietic stem cell mobilization

“…2,[6][7][8] Osteoblasts, in particular, have been implicated in controlling HSC numbers, and studies in gene-targeted 2 and hormone-treated 6,9 mice show a strong correlation between experimentally induced expansion of osteoblasts and increased HSC frequency. Significantly, most studies of osteoblast function as it relates to HSC have relied on complex in vivo models [10][11][12][13] or on in vitro systems in which osteoblasts are derived ex vivo by extended culture of calvarial precursor cells. 10 Although clearly suggestive, these in vivo analyses are complicated by the unavoidable presence of other, nonosteoblastic cell types, whereas in vitro studies of culture-derived osteoblasts are challenged by the possibility that extended culture may induce changes in osteoblast behavior and/or may fail to properly recapitulate the in vivo conditions under which osteoblasts normally would be formed or regulated.…”

Thrombin, but not bradykinin, stimulates proliferation in isolated human osteoblasts, via a mechanism not dependent on endogenous prostaglandin formation

“…As it is not clear whether one can extrapolate results from rat or murine cell lines to non-transformed human bone cells, we have used primary cultures of normal human osteoblast-like cells in this report. This is currently believed to be the most reliable cell culture system since it reflects processes occumng in human bone and we have recently optimized an isolation technique providing cells with a mature osteoblastic phenotype (Jonsson et al 1999). Using these cells, we here demonstrate that thrombin stimulates proliferation in non-transformed human osteoblasts and that the mechanism appears to involve the tetheric G-protein coupled thrombin receptor.…”

Role of DNA methylation in the regulation of the RANKL-OPG system in human bone

“…Trabecular bone cylinders of the central part of the femoral head (thus avoiding the fractured and the subchondral regions) were obtained with a trephine, washed extensively in phosphate-buffered saline, snap-frozen in liquid nitrogen and stored at -80°C or used to generate primary human osteoblasts (hOBs) by the explant technique. 40 The osteoblastic phenotype was confirmed by the ability to express osteocalcin when stimulated with vitamin D and to form a mineralized matrix. hOBs were cultured in DMEM supplemented with 10% FBS and antibiotics.…”