DMEM(低糖，含双抗) 细胞培养

DMEM是一种含各种氨基酸和葡萄糖的培养基，高糖型有利于细胞停泊于一个位置生长，适于生长较快、附着较困难肿瘤细胞等。该类培养基广泛应用于疫苗生产和各种初代病毒宿主细胞的细胞培养及单一细胞培养;如A9、3T6、BALB/3T3、COS-1、COS-3、COS-7、L6、WEHI-3b等细胞系的培养。

DMEM培养基的配制方法：

1 制备新鲜三蒸水或Millipore超纯水。

2 称取所需量的干粉培养基，加入约终体积一半的三蒸水中;若配制一个包装的培养液，在将整个包装的干粉倒入三蒸水后，需用水洗包装袋内面2次，倒入培养液中，以保证所有干粉都溶解成培养液。磁力搅拌或人工搅拌使之*溶解。

3 根据包装袋上的要求补加所需量的碳酸氢钠;根据实验需要，添HEPES(5-20mmol/L)、谷氨酰胺和其他特殊物质。

4 加水定容到终体积。

5 必要时用1 mol/L 盐酸和1 mol/L 氢氧化钠调节pH。

6 用无菌0.22um滤膜过滤除菌，分装于无菌血清瓶中，4℃冰箱保存。

配制好的培养液用前加入100U/mL青霉素和100U/mL链霉素，并根据需要加入血清(5%-20%)。

DMEM培养基的特点：

（1）氨基酸含量为依格尔培养基的2倍，且含有非必需氨基酸，如甘氨酸等；

（2）维生素含量为依格尔培养基的4倍；

（3）含有糖酵解途径中的重要物质——丙酮酸；

（4）含有微量的铁离子。

DMEM培养基应用广泛。基于DMEM培养基中氨基酸、微生物含量丰富，并具有糖酵解中的丙酮酸和微量Fe离子，因此在细胞培养、疫苗生产等方面的应用为常见。

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DMEM(低糖，含双抗) 细胞培养