Table I.
Summary of the optimal conditions for culturing bone-forming rodent osteoblasts.
| Conditions | Rat | Mouse |
|---|---|---|
| Cell isolation protocol | TCC | TCC or CCEC |
| Cell yield/calvariae (after expansion for 2–3 days) | 7×106–107 | 3–5×106 |
| Basal medium | DMEM or αMEM Cultures 3–4 days shorter in αMEM |
αMEM |
| FCS | 10% FCS or HI FCS | 10% FCS or HI-FCS |
| Dexamethasone | 10 nM | Not required |
| β-glycerophosphate | 2 mM | 2–5mM |
| Ascorbate | 50 μg/ml | 50 μg/ml |
| Plate format | 6-, 12- or 24-well plates | 6-well plates |
| Culture duration | 14–17 days in DMEM 10–14 days in αMEM |
21–28 days |
TCC, trypsin-collagenase-collagenase; CCEC, collagenase-collagenase-EDTA-collagenase.