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當(dāng)前位置：首頁 > 技術(shù)文章 > 細(xì)胞培養(yǎng)中培養(yǎng)基的作用

細(xì)胞培養(yǎng)中培養(yǎng)基的作用

2022-01-13 瀏覽次數(shù)：1769

1.液體培養(yǎng)基貯存于4℃ 冰箱，避免光照，實(shí)驗(yàn)進(jìn)行前放在37℃ 水槽中溫?zé)帷?S'、


2.液體培養(yǎng)基(加血清)存放期為六個(gè)月，期間glutamine可能會(huì)分解，若細(xì)胞生長不佳
可以再添加適量glutamine。
3.粉末培養(yǎng)基配制(以1升為例)：
3.1.細(xì)胞培養(yǎng)基通常須添加10%血清，因此粉末培養(yǎng)基之配制體積為900ml，pH為7.2-7.4。NaHCO3為另外添加，若將NaHCO3粉末直接加入液體培養(yǎng)基中會(huì)造成pH之誤差，或局部過堿。因此粉末培養(yǎng)基及NaHCO3粉末應(yīng)分別溶解后才混合，然后用CO2氣體調(diào)整pH，而非用強(qiáng)酸(HCl)或強(qiáng)堿(NaOH)，因?yàn)槁入x子對(duì)細(xì)胞生長可能有影響，且貯存時(shí)培養(yǎng)基的pH易發(fā)生改變。生物3.2.材料：純水(milli-Q水或二次至三次蒸餾水，水品質(zhì)非常重要)，粉末培養(yǎng)基，NaHCO3
電磁攪拌器,無菌血清瓶,0.1或0.2mm無菌過濾膜,pH計(jì)，真空泵，CO2氣體。
3.3.步驟：
3.3.1.取粉末培養(yǎng)基溶于700mlmilli-Q水中，攪拌使其溶解。
3.3.2.稱取適量之NaHCO3粉末溶于200mlmilli-Q水中，攪拌使其溶解，然后通入CO2氣體至飽和，約3-5分鐘。
3.3.3.將溶解且含飽和CO2之NaHCO3溶液加入溶解之液體培養(yǎng)基中混合?；旌笕芤褐畃H應(yīng)為7.2-7.4，除非pH值偏差太大，否則不需用酸堿再調(diào)整之。若為太堿，可再通入CO2氣體調(diào)整pH。培養(yǎng)基以真空幫浦通過過濾膜時(shí)，pH會(huì)升高0.1-0.2。
3.3.4.以0.1或0.2mm無菌過濾膜過濾滅菌，同時(shí)分裝至無菌容器中，標(biāo)示培養(yǎng)基種類、日期、瓶號(hào)等，貯存于4℃。(血清亦可加入培養(yǎng)基中一起過濾)
3.3.5.配制之培養(yǎng)基配制須作生長試驗(yàn)與污染測(cè)試。



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