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细胞培养常规方法

浏览次数:509发布日期:2020-08-27

细胞培养常规方法

 

 

1. 冻存细胞的复苏

  1. YINGZUNSHOUMANDONGKUAIRONGDEYUANZE。XIANJIANGSHUIYUGUODIAOZHI37-37。5DU,QUCHUDONGCUNDEXIBAOXUNSUFANGRUHOUJIANGXIBAOMIANJINZHISHUIMIANYIXIABUDUANYAODONGZHIRONGHUA。

  2. ZAIWUJUNTAINEIJIANGWANQUANPEIYANGJIJIARU50mlDEXIAOPEIYANGPINGNEI,YUE5mlZUOYOU,RANHOUYONGWUJUNXIGUANCONGDONGCUNGUANNEIQUCHUXIBAO,ZHIPEIYANGBINGNEIQINGQINGYAOHUANG,SHIXIBAOJUNYUNHOUZHIPEIYANGXIANGNEIPEIYANG。

2. 传代:

DUIYUTIEBIXIBAOYINGXIANXI(DAO)JINPEIYANGJI,XIDEYUEGANJINGYUEHAO,YIMIANZHONGHEHOUJIARUDEXIAOHUAYE,SHIQIANGDUJIANRUO(HUOPBSXI1-3CI)。50mlPEIYANGPINGJIARUXIAOHUAYEYUE1-3ml,ANCIBILIJINXINGXIAOHUA,(GENJUJINGYAN),HUANGDONGSHIXIAOHUAYEPUJUNYUNZHI37DUPEIYANGXIANGYUE2-5FENZHONG,JINGXIAJIANXIBAOSHOUSUOBIANYUANHUOSHAOSHUTUOLUOHOU,QINGQINGZHENDONGPINGDISHIXIBAOQUANBUTUOLUO,JIARU2-3mlWANQUANPEIYANGJIHOU,QINGQINGCHUIDA,SHIXIBAOJIBENCHENGDANGEXUANFU,RANHOUFENZHIQITAWUJUNPEIYANGPINGNEI,JIARUWANQUANPEIYANGJIHOUJIXUPEIYANGHUOSHIYAN。

YIBANCHUANDAIKEZHIJIEJIANGXIBAOYUANYEFENZHIQITAPEIYANGPINGNEI,JIARUWANQUANPEIYANGJIJIXUPEIYANG,RUYAOGAONONGDUKEXIANLIXIN1000rpm,5minHOUJIARUWANQUANPEIYANGJI,QINGQINGCHUIYUNHOU,FENZHIQITAPEIYANGPINGNEIJIARUWANQUANPEIYANGJIJIXUPEIYANG。

  1. TIEBIXIBAO:

  2. XUANFUXIBAO:

3. 冻存 

将贴壁细胞消化后离心收集,悬浮细胞直接离心收集,以完全培养基或胎牛血清重悬细胞至终浓度约106/ml。加入10%的DMSO。以每管1~2ml分装至冻存管中。用绝热材料包裹置-70摄氏度冰箱冷冻过夜。次日保存到液氮中。

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