汉恒悬浮细胞专用腺病毒特点
1.XUANFUXIBAOZHUANYONG
HANHENGSHENGWUKAIFADEXUANFUXIBAOZHUANYONGXIANBINGDUZHENDUIXUANFUXIBAODEXIBAOMOTEXINGJINXINGLEXIANBINGDUDEGAIZAO，BINGDUGANRANTIXIYEJINXINGLEXIANGYINGDEYOUHUA，JIDAGAISHANDUIXUANFUXIBAODEGANRANNENGLI。DUITIEBIXIBAODEGANRANNENGLIWUYINGXIANG，KEYITONGYONG。
2. CHAOQIANGGANRANXIAOLV
HANHENGXUANFUXIBAOZHUANYONGXIANBINGDUDUIJurkat、K562DENGZHONGDUOXUANFUPEIYANGXIBAOXIGANRANXIAOLVJIEJIN100%，DUIXUEYELAIYUANYUANDAIXIBAO，RUCD4+ TXIBAOGANRANXIAOLVKEDA60%YISHANG。
腺病毒的细胞感染特性
1）XIANBINGDUBUCHARUXIBAOJIYINZU，BUXINGCHENGWENDINGZHUANRAN；
2）XIANBINGDUDESHUNSHIBIAODANENGLIYIBANWEI2-4ZHOU，XIBAOZENGZHIYUEKUAI，BIAODASHIJIANYUEDUAN；
3）YOUYUBUHUISUIJICHARUSUNSHANGJIYINZU，XIANGBIJIAOMANBINGDU，XIANBINGDUDUITIWAIPEIYANGDEMUDEXIBAODUXINGJIAOXIAO，YINCIKEYIYONGJIAOGAOMOILAIGANRANMUDEXIBAO。
4）GAODIDU：HANHENGSHENGWUTIGONGDEXIANBINGDUYABOappGUANWANGDIDUJIAOGAO，YOUYUCAIYONGLEHANHENGTEYOUDETIXI，YONGYUXIBAOGANRANJIBIEDEXIANBINGDUYIBANKEYIDADAO1010 PFU/mlDIDU。
腺病毒细胞实验策略
YOUYUXIANBINGDUDEFEIWENZHUANXING、DILITIXIBAODUXINGYIJIGAODIDU，YONGXIANBINGDUGANRANXIBAODEYIBANCELVESHI：XIANKUOZENGZULIANGDEMUDEXIBAO，RANHOUYONGXIANBINGDUGANRANMUDEXIBAO，DAIMUDEJIYINBIAODAHOUZHIJIEJINXINGHOUXUDEXIBAOGONGNENGXINGSHIYAN。YIBANBUHUIYONGXIANBINGDUXIANGANRANXIBAOHOUZAIDUIXIBAOJINXINGERCIKUOZENGFANGDAHUAPEIYANG。
悬浮细胞专用腺病毒感染实验相关知识点
1）XUANFUXIBAOZHUNBEI：XIANBINGDUJIANWENZHUANXIBAOXIQIAN，QINGQUEBAOMUDEXIBAOZHUANGTAILIANGHAO、WUZHIYUANTIWURAN。XIANBINGDUGANRANDESHIHOUXIBAOHUIHELVWEI40%～60%JIANWEIYI。
2）GANRANXIBAOZUIJIAMOIDECEDING：MOI（GANRANFUSHU）SHIZHIMEIGEXIBAOGANRANDEBINGDUSHU。
ZHU：XIANBINGDUDUIYUBUTONGZHONGLEIBUTONGLAIYUANDEXIBAO，QIZUISHIMOIGEYOUCHABIE。DUIYUXUANFUXIBAO，XIANBINGDUMOIYIBANYI30, 100, 300, 1000, 2000DEBILIDIZENGJINXINGTIDUMOSUO，XIANGBIYUTIEBIXIBAO，XUANFUXIBAODEXIANBINGDUMOIXUQIUGENGGAO，MOITIDUMOSUOJIANYIYONG96KONGBANJINXINGYIJIESHENGBINGDUHEXIBAO。
3）MOIDUIYINGBINGDUTIJIDEHUANSUAN（FEICHANGZHONGYAO!）
感染时细胞数（一般以传代计数细胞数×2计算）×MOI=病毒个数；则加入的病毒体积数=病毒个数/病毒滴度
比如第一天传代1×104细胞到96孔板一个孔，第二天长到2×104细胞，按照MOI=500计算，需加入病毒总量为2×104×500=1×107个病毒粒子，按照腺病毒1×1010 PFU/ml的滴度，则加入的病毒体积数为(1×107)/(1×1010 PFU/ml)=1×10-3 ml=1 μl。
腺病毒感染细胞步骤
1.GANRANQIANZHUNBEI
1）ANSHIYANXUYAOJIANGXUANFUXIBAOPUBAN（BIRU96KONGBAN）37 ℃ PEIYANGGUOYE。XIBAOSHUYIDI2TIANMIDUYUE40%～60%JIANWEIYI。
2）CONGBINGXIANGQUCHUXUANFUXIBAOZHUANYONGXIANBINGDUBINGZAIBINGSHANGHUANXIANRONGHUA，DAIWANQUANRONGHUAHOUKAISHIBINGDUGANRANSHIYAN。
2. MUDEXIBAODEGANRAN
XIQUXIBAOYUANYOUPEIYANGJI，JIARUXINXIANPEIYANGJI，XUANDINGMOIZHIHUANSUANDUIYINGDEBINGDUYUANYETIJI，BINGANTIJIJIANGBINGDUYUANYEJIARUXIBAOZHONG，YAOYUN。FENGHAOKOU，FANGRUPINGJIAOLIXINJIHOU，DISULIXIN1.5h，RANHOUFANGRU37℃PEIYANGXIANGZHONGJIXUJIGANRAN4XIAOSHI。
1) 37 ℃PEIYANGXIANGZHONGGANRANWANBIHOU，QUCHUXIBAO，XIQUHANBINGDUDEPEIYANGYE，HUANSHANGXINXIANPEIYANGJI，JIXU37℃PEIYANG
2)GANRAN48XIAOSHIHOU，YINGGUANGXIANWEIJINGJIANCEGFP/RFPDENGYINGGUANGBIAODAXIAOLV。DUIYUBUFENXUANFUXIBAO，YINGGUANGBIAODAJIAOMAN，XUDAI96XIAOSHIHOUCAIHUIYOUMINGXIANYINGGUANG
ZHU：WEIXINSAOMAGUANZHUHANHENGSHENGWUGONGZHONGHAO，GUANKANHANHENGSHENGWU-XIANBINGDUXIBAOGANRANSHIYANCAOZUOJIAOXUESHIPIN
悬浮细胞专用病毒感染实例
病毒：悬浮细胞专用腺病毒Ads-GFP，1×1010 PFU/ml
感染细胞：Jurkat、K562、HL-60、L2110
感染方式：MOI=500
检测方法：感染72H后荧光显微镜检测
悬浮细胞专用腺病毒Ads感染小鼠原代CD4+ T细胞
XIBAO：XIAOSHUYUANDAICD4+ TXIBAO
CIJIFANGSHI：CD3/CD28KANGTIHEIL-2JIHUO48h
BINGDU：Ads-GFP, 1×10^10 PFU/ml
SHIYONGFANGSHI：MOI=500
悬浮细胞专用腺病毒Ads感染人原代CD4+ T细胞