Briefly, cancellous bone was disintegrated mechanically and washed 3–5 times with Dulbecco’s phosphate buffered saline (DPBS). After 1 h of collagenase digestion (DPBS, 0.07% collagenase II—Biochrom AG, Berlin, Germany) at 37 °C, cancellous bone pieces were washed with DPBS and released OBs were transferred to cell culture flasks in growth medium (MEM/Ham’s F12, 10% FCS, 2 mM l -glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin, 50 μM l -ascorbate-2-phosphate, 50 μM β-glycerol phosphate) for expansion. Medium was changed every 3–4 days [22 (link),23 (link)].
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