American Society of Hematology

Figure 3.

$Evaluation of osteoclast progenitor populations in the CD45+ fraction from EB cultures. (A) Four discrete populations defined by their expression of CD11b and CD14 were sorted to purity and plated in triplicate at equal numbers (4 K). They were then cultured in M-CSF and RANKL for 14 days. (B-C) TRAcP+ cells generated from each population were photographed at ×14 and ×35 magnification and enumerated. Most of the osteoclastogenic activity was contained in the CD11bCD14 DN population. (D) The DN population was further dissected with CD15 and sorted into CD15+ and CD15−/low. The latter population contained a small population expressing CD115. These populations were sorted and plated at equal numbers into (E-F) osteoclast inducing media and (G) myeloid-erythroid methylcellulose for 10 days. TRAcP+ osteoclasts and colonies were enumerated. TRAcP-stained wells were imaged using a LeciaEZ4D stereomicroscope.$

Evaluation of osteoclast progenitor populations in the CD45⁺ fraction from EB cultures. (A) Four discrete populations defined by their expression of CD11b and CD14 were sorted to purity and plated in triplicate at equal numbers (4 K). They were then cultured in M-CSF and RANKL for 14 days. (B-C) TRAcP⁺ cells generated from each population were photographed at ×14 and ×35 magnification and enumerated. Most of the osteoclastogenic activity was contained in the CD11bCD14 DN population. (D) The DN population was further dissected with CD15 and sorted into CD15⁺ and CD15^−/low. The latter population contained a small population expressing CD115. These populations were sorted and plated at equal numbers into (E-F) osteoclast inducing media and (G) myeloid-erythroid methylcellulose for 10 days. TRAcP⁺ osteoclasts and colonies were enumerated. TRAcP-stained wells were imaged using a LeciaEZ4D stereomicroscope.

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