二価鉄の培養3T3細胞の生存率に及ぼす影響

細胞培養レベルでの二価鉄の影響をマウス3T3細胞を用いてコロニー法でみた結果は、細胞播種直後の二価鉄添加（濃度0.5mMで0.75％の生存率）と培地に二価鉄添加後の細胞播種（濃度0.5mMで55％の生存率）で異なった細胞生存率が得られたことから、いずれも単細胞処理後の浮遊細胞に対する影響と考慮して、ディシュ面への接着後の培養細胞に対する二価鉄の影響を検討した。細胞播種1日後の二価鉄添加では濃度0.5mMで73％の生存率が得られ、上記条件下よりも高率の生存率が認められた。さらに生存の細胞コロニーの径は処理群で短縮がみられ、増殖阻害が明らかに認められた。細胞播種直後の二価鉄添加の生存率曲線から得られた損傷回復が影響されない最低濃度0.25mMでの二価鉄の時間的処理では、1時間処理では統計的に有意差は認められなかったが、3時間、6時間処理で有意差の増殖阻害が認められた。

Previous reports demonstrated that adding ferrous iron into the culture medium immediately after seeding of cells showed significantly cell killing activity (0.75% survival by 0.5mM ferrous iron) than the case of the cell seeding after adding ferrous iron. In this report, the effect of an addition of ferrous iron after the adhesion of cells onto the dish, in the condition 1 day after cell seeding, was investigated by the colony method. The dose responsed survival curve obtained above condition showed nealy survival curve in the case of the cell seeding after addition of ferrous iron, and 72.9% survival by 0.5mM ferrou iron was estimated. But the colony diameter was shortened depending on the Fe(2+) doses, the 0.5mM-affected colony showed a half of the diameter of control colony, namely proliferation of the cells was inhibited. The 1 hour treatment with ferrous iron (0.25mM) did not inhibited, but both the 3 and 6 hours treatments inhibited significantly a proliferation of the cells checked by the diameter of colony. The facts suggest that both the oxidation mechanism of ferrous iron, induced radicals, promptly occured at the time of an addition of ferrous iron into the culture medium and the followed oxido-reductive reactions are concerned in the injury of cells.