DMEM（H）（含雙抗，不含丙酮酸）細胞培養

DMEM（H）（含雙抗，不含丙酮酸）細胞培養

DMEM是一種含各種氨基酸和葡萄糖的培養基，高糖型有利于細胞停泊于一個位置生長，適于生長較快、附著較困難腫瘤細胞等。該類培養基廣泛應用于疫苗生產和各種初代病毒宿主細胞的細胞培養及單一細胞培養;如A9、3T6、BALB/3T3、COS-1、COS-3、COS-7、L6、WEHI-3b等細胞系的培養。

DMEM培養基的成分

DMEM培養基的配制方法：

1 制備新鮮三蒸水或Millipore超純水。

2 稱取所需量的干粉培養基，加入約終體積一半的三蒸水中;若配制一個包裝的培養液，在將整個包裝的干粉倒入三蒸水后，需用水洗包裝袋內面2次，倒入培養液中，以保證所有干粉都溶解成培養液。磁力攪拌或人工攪拌使之*溶解。

3 根據包裝袋上的要求補加所需量的碳酸氫鈉;根據實驗需要，添HEPES(5-20mmol/L)、谷氨酰胺和其他特殊物質。

4 加水定容到終體積。

5 必要時用1 mol/L 鹽酸和1 mol/L 氫氧化鈉調節pH。

6 用無菌0.22um濾膜過濾除菌，分裝于無菌血清瓶中，4℃冰箱保存。

配制好的培養液用前加入100U/mL青霉素和100U/mL鏈霉素，并根據需要加入血清(5%-20%)。

DMEM培養基的特點：

（1）氨基酸含量為依格爾培養基的2倍，且含有非必需氨基酸，如甘氨酸等；

（2）維生素含量為依格爾培養基的4倍；

（3）含有糖酵解途徑中的重要物質——丙酮酸；

（4）含有微量的鐵離子。

DMEM培養基應用廣泛。基于DMEM培養基中氨基酸、微生物含量豐富，并具有糖酵解中的丙酮酸和微量Fe離子，因此在細胞培養、疫苗生產等方面的應用為常見。

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