细胞培养常规方法

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。