I know that extracellular calcium can replenish intracellular calcium stores. However, they myriad of papers on the issue appear to be contradictory as to the requirement for extracellular calcium in phagocytic cells.
Methodological differences may have given rise to discrepancies.
If you are interested in extracellular calcium, then perhaps calcium free buffer / medium would be better choice for experimental design.
You can explore a design involving first deprivation, then replenishment to see if there's a rescue effect.
Although EGTA has higher affinity for calcium than for magnesium, its spillover effects cannot be ruled out in its presence.
https://en.wikipedia.org/wiki/EGTA_(chemical)
It's better to have more than one method to confirm things.
Good luck.
Lance,
Not sure why you need to maintain macrophages in EGTA for 12 hrs and whether this would be at 37° or 4°C. If at 37° C, then it is highly likely that cell injury will occur since cell metabolism, respiration and ATPase function will likely be inhibited in the absence of calcium and magnesium (as Dr. Wong mentioned, EGTA is not specific for calcium). If at 4°C, then this treatment will probably have much less of an effect on macrophage function, but some effects are still possible.
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