Osteoclast generation from human fetal bone marrow in cocultures with murine fetal long bones

Abstract

Osteoclast formation in vitro from human progenitor cells was studied in cocultures of periosteum-free murine long-bone rudiments and human fetal tissues. No osteoclasts were generated from chorionic villi or from fetal liver, but bone marrow and purified bone-marrow fractions gave rise to multinucleated cells that resorbed calcified cartilage matrix. These polykarya react very strongly for tartrate-resistant acid phosphatase (TrAP) and upon ultrastructural examination show large ruffled borders in areas of resorption. Resorption of murine calcified cartilage matrix by human osteoclasts was less than resorption. by osteoclasts formed from murine fetal bone-marrow cells. Our results show that the murine long-bone rudiment can be used to generate osteoclasts from human sources of progenitor cells and to assess the biological activity of the formed osteoclasts. This coculture system thereby offers possibilities to study human osteoclast pathology in vitro. The use of TrAP as marker for osteoclasts in cell cultures is discussed.

The Keap1/Nrf2 Protein Axis Plays a Role in Osteoclast Differentiation by Regulating Intracellular Reactive Oxygen Species Signaling

“…The PCR primers used in the experiments were from Primerbank and are described in Table 1. We chose six representative marker genes (ATP6v0d2 (26), cathepsin K (27), dendritic cell-specific transmembrane protein (DC-STAMP) (28), matrix metallopeptidase-9 (MMP9) (29), osteoclast-associated receptor (OSCAR) (30), and TRAP (31) for osteoclast differentiation.…”

The Role of Inflammatory Cytokines, the RANKL/OPG Axis, and the Immunoskeletal Interface in Physiological Bone Turnover and Osteoporosis

“…Binding of RANKL to RANK initiates a sequence of signal transduction pathways mediated though TNF receptor-associated factor 6 (TRAF6) and including NF- κ B [38–40], c-Jun N-terminal kinase (JNK)/cJun/fos [38, 41–43], and nuclear factor of activated T cells (NFAT) [44–46] that initiate the differentiation of the early osteoclast precursor into a preosteoclast (recognized in vitro or ex vivo by the expression of the enzyme tartrate resistant acid phosphatase (TRAP)). Preosteoclasts ultimately fuse with each other into mature multinucleated bone resorbing osteoclasts recognized by the expression of key osteoclast markers including TRAP [47], calcitonin receptors [28], cathepsin K [48], pp60c-src [49], matrix metalloproteinase 9 (MMP9) [50], and the alpha V beta 3 integrin chains [51, 52]. Osteoclasts originating from multiple species can further be identified by reactivity to a unique osteoclast-specific monoclonal antibody (121F) [53].…”

Postmenopausal Osteoporosis: The Role of Immune System Cells

“…Mature multinucleated bone resorbing OCs are recognized by the expression of key OC markers including TRAP [18], calcitonin receptors [19], cathepsin K [20], pp60c-src [21], matrix metalloproteinase 9 (MMP9) [22], and the alpha V beta 3 integrin chains [23, 24]. …”