[求救] 細胞RAW264.7培養繼代問題
我想請問養過這株RAW264.7的細胞株嗎?
我目前正在做細胞的繼代
但是Passage2~3次後
隔天去看細胞幾乎都活化了
細胞來源:BCRC
培養基:90% GIBCO DMEM 的 Power with 4mM L-glutamine,
4500mg/L Glucose 另加1.5g Sodium bicarbonate
10% GIBCO 的 FBS
繼代方法: 1.滿盤時,吸除舊的medium,加入1ml新鮮medium至25T來刮細胞
2.吸取 1ml細胞溶液和9ml新鮮medium在15ml離心管均勻pipette
3.以1拆5去分盤
我的問題有以下4個
1.Passage2~3次後,隔天去看細胞幾乎全部都活化了
2.是否是繼代方法錯誤?還是中間步驟有需要改進的地方?
3.是否是medium中L-glutamine沒有再補充的原因?
(原本medium中L-glutamine的濃度是足夠的)
4.是否是FBS的品質不夠好,還是Medium要買Liquid的?
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