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https://hdl.handle.net/2440/38771
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Type: | Journal article |
Title: | Cloning and characterization of osteoclast precursors from the raw264.7 cell line |
Author: | Cuetara, B. Crotti, T. O'Donoghue, A. McHugh, K. |
Citation: | In Vitro Cellular and Developmental Biology: Animal, 2006; 42(7):182-188 |
Publisher: | Soc in Vitro Biology |
Issue Date: | 2006 |
ISSN: | 1071-2690 1543-706X |
Abstract: | Osteoclasts are bone-resorbing cells that differentiate from macrophage precursors in response to receptor activator of NF-kappaB ligand (RANKL). In vitro models of osteoclast differentiation are principally based on primary cell cultures, which are poorly suited to molecular and transgene studies because of the limitations associated with the use of primary macrophage. RAW264.7 is a transfectable macrophage cell line with the capacity to form osteoclast-like cells. In the present study, we have identified osteoclast precursors among clones of RAW264.7 cells. RAW264.7 cell were cloned by limiting dilution and induced to osteoclast differentiation by treatment with recombinant RANKL. Individual RAW264.7 cell clones formed tartrate resistant acid phosphatase (TRAP)-positive multinuclear cells to various degrees with RANKL treatment. All clones tested expressed the RANKL receptor RANK. Each of the clones expressed the osteoclast marker genes TRAP and cathepsin-K mRNA with RANKL treatment. However, we noted that only select clones were able to form large, well-spread, TRAP-positive multinuclear cells. Clones capable of forming large TRAP-positive multinuclear cells also expressed beta3 integrin and calcitonin receptor mRNAs and were capable of resorbing a mineralized matrix. All clones tested activated NF-kappaB with RANKL treatment. cDNA expression profiling of osteoclast precursor RAW264.7 cell clones demonstrates appropriate expression of a large number of genes before and after osteoclastic differentiation. These osteoclast precursor RAW264.7 cell clones provide a valuable model for dissecting the cellular and molecular regulation of osteoclast differentiation and activation. |
Keywords: | Cell Line Clone Cells Osteoclasts Stem Cells Animals Mice Bone Resorption Durapatite Acid Phosphatase Isoenzymes Carrier Proteins Membrane Glycoproteins Receptors, Calcitonin Integrin beta Chains Recombinant Fusion Proteins DNA, Complementary RNA, Messenger Genetic Markers Gene Expression Profiling Cell Differentiation RANK Ligand Receptor Activator of Nuclear Factor-kappa B Tartrate-Resistant Acid Phosphatase |
DOI: | 10.1290/0510075 |
Published version: | http://dx.doi.org/10.1290/0510075.1 |
Appears in Collections: | Aurora harvest Pathology publications |
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