DMEM（H）（含雙抗，不含丙酮酸）細(xì)胞培養(yǎng)

DMEM（H）（含雙抗，不含丙酮酸）細(xì)胞培養(yǎng)

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

DMEM培養(yǎng)基的成分

DMEM培養(yǎng)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(yǎng)基，加入約終體積一半的三蒸水中;若配制一個(gè)包裝的培養(yǎng)液，在將整個(gè)包裝的干粉倒入三蒸水后，需用水洗包裝袋內(nèi)面2次，倒入培養(yǎng)液中，以保證所有干粉都溶解成培養(yǎng)液。磁力攪拌或人工攪拌使之*溶解。

3 根據(jù)包裝袋上的要求補(bǔ)加所需量的碳酸氫鈉;根據(jù)實(shí)驗(yàn)需要，添HEPES(5-20mmol/L)、谷氨酰胺和其他特殊物質(zhì)。

4 加水定容到終體積。

5 必要時(shí)用1 mol/L 鹽酸和1 mol/L 氫氧化鈉調(diào)節(jié)pH。

6 用無(wú)菌0.22um濾膜過濾除菌，分裝于無(wú)菌血清瓶中，4℃冰箱保存。

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

DMEM培養(yǎng)基的特點(diǎn)：

（1）氨基酸含量為依格爾培養(yǎng)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(yǎng)基的4倍；

（3）含有糖酵解途徑中的重要物質(zhì)——丙酮酸；

（4）含有微量的鐵離子。

DMEM培養(yǎng)基應(yīng)用廣泛。基于DMEM培養(yǎng)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(xì)胞培養(yǎng)、疫苗生產(chǎn)等方面的應(yīng)用為常見。

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