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PT67小鼠逆轉(zhuǎn)錄病毒包裝細(xì)胞細(xì)胞培養(yǎng)方法 
1、收到細(xì)胞，請(qǐng)查看瓶子是否有破裂，培養(yǎng)基是否漏出，是否渾濁，如有請(qǐng)盡快。
2、收到細(xì)胞，如包裝完好，請(qǐng)?jiān)陲@微鏡下觀察細(xì)胞。,由于運(yùn)輸過(guò)程中的問(wèn)題，細(xì)胞培養(yǎng)瓶中的貼壁細(xì)胞有可能從瓶壁中脫落下來(lái)，顯微鏡下觀察會(huì)出現(xiàn)細(xì)胞懸浮的情況，出現(xiàn)此狀態(tài)時(shí)，請(qǐng)不要打開(kāi)細(xì)胞培養(yǎng)瓶，應(yīng)立即將培養(yǎng)瓶置于細(xì)胞培養(yǎng)箱里靜止3-5小時(shí)左右，讓細(xì)胞先穩(wěn)定下，再于顯微鏡下觀察，此時(shí)多數(shù)細(xì)胞會(huì)重新貼附于瓶壁。如細(xì)胞仍不能貼壁，請(qǐng)用臺(tái)盼藍(lán)染色法鑒定細(xì)胞活力，如臺(tái)盼藍(lán)染色證實(shí)細(xì)胞活力正常請(qǐng)按懸浮細(xì)胞的方法處理
3. 收到細(xì)胞時(shí)如無(wú)異常情況 ，請(qǐng)?jiān)陲@微鏡下觀察細(xì)胞密度，如為貼壁細(xì)胞，未超過(guò)80%匯合度時(shí)，將培養(yǎng)瓶中培養(yǎng)基吸出，留下 5-10ML培養(yǎng)基繼續(xù)培養(yǎng)：超過(guò)80%匯合度時(shí)，請(qǐng)按細(xì)胞培養(yǎng)條件傳代培養(yǎng)。如為懸浮細(xì)胞，吸出培養(yǎng)液，1000轉(zhuǎn)/分鐘離心3分鐘，吸出上清，管底細(xì)胞用新鮮培養(yǎng)基懸浮細(xì)胞后移回培養(yǎng)瓶。
4，將培養(yǎng)瓶置于37℃培養(yǎng)箱中培養(yǎng)，蓋子微微擰松。吸出的培養(yǎng)基可以保存在滅菌過(guò)的瓶子里，存放于4℃冰箱，以備不時(shí)之需。
5 、24小時(shí)后，細(xì)胞形態(tài)已恢復(fù)并貼滿瓶壁，就可以傳代了。（貼壁細(xì)胞）將培養(yǎng)瓶里的培養(yǎng)基倒去，加3-5ml（以能覆蓋細(xì)胞生長(zhǎng)面為準(zhǔn)）PBS或Hanks’液洗滌后棄去。加0.5-1ml  0.25%含EDTA的胰酶消化，消化時(shí)間以具體細(xì)胞為準(zhǔn)，一般1-3分鐘，不超過(guò)5分鐘。可以放入37℃培養(yǎng)箱消化。輕輕晃動(dòng)瓶壁，見(jiàn)細(xì)胞脫落下來(lái)，加入3-5ml培養(yǎng)基終止消化。用移液管輕輕吹打瓶壁上的細(xì)胞，使之*脫落，然后將溶液吸入離心管內(nèi)離心，1000rpm/5min。棄上清，視細(xì)胞數(shù)量決定分瓶數(shù)，一般一傳二，如細(xì)胞量多可一傳三，有些細(xì)胞不易傳得過(guò)稀，有些生長(zhǎng)較快的細(xì)胞則可以多傳幾瓶，以具體細(xì)胞和經(jīng)驗(yàn)為準(zhǔn)。（懸浮細(xì)胞）用移液管輕輕吹打瓶壁，直接將溶液吸入離心管離心即可。
6，貼壁細(xì)胞 ，懸浮細(xì)胞。嚴(yán)格無(wú)菌操作。換液時(shí)，換新的細(xì)胞培養(yǎng)瓶和換新鮮的培養(yǎng)液，37度    5%CO2 培養(yǎng)
7.  收到細(xì)胞后，請(qǐng)鏡下觀察細(xì)胞，用恰當(dāng)方式處理細(xì)胞。若懸浮的細(xì)胞較多，請(qǐng)離心收集細(xì)胞，接種到一個(gè)新的培養(yǎng)瓶中。棄掉原液，使用新鮮配制的培養(yǎng)基，使用進(jìn)口胎牛血清. 剛接到細(xì)胞，若細(xì)胞不多時(shí) 血清濃度可以加到15％去培養(yǎng)。若細(xì)胞迏到80%左右 ，血清濃度還是在10％。
培養(yǎng)基參考
500ml基礎(chǔ)培養(yǎng)基（含：必需與非必需氨基酸、維他命、有機(jī)與無(wú)機(jī)成分、激素、生長(zhǎng)因子與微量元素）、細(xì)胞生長(zhǎng)添加劑、抗生素/抗菌素溶液或青霉素/鏈霉素（P/S）溶液、FBS，適合在飽和濕度為5%CO2與95%空氣的培養(yǎng)箱中使用
菅囊酵母ATCC32691    QT-00115
弗氏檸檬酸桿菌ATCC10787    QT-00116
莢膜紅細(xì)菌ATCC11166    QT-00117
發(fā)根土壤桿菌（發(fā)根植物單胞菌）ATCC11325    QT-00118
生孢梭菌（產(chǎn)芽孢梭狀芽孢桿菌）ATCC11437    QT-00119
痤瘡丙酸桿菌ATCC11827    QT-00120
地衣芽孢桿菌ATCC11946    QT-00121
谷氨酸棒桿菌ATCC13032    QT-00122
熏衣草灰鏈霉菌ATCC13306    QT-00123
豬霍亂沙門氏菌豬霍亂亞種ATCC13312    QT-00124
脫氮假單胞菌ATCC13867    QT-00125
耐放射異常球菌ATCC13939    QT-00126
綠針假單胞菌ATCC13985    QT-00127
腸炎沙門氏菌腸炎亞種ATCC14028    QT-00128
巨大芽孢桿菌ATCC14581    QT-00129
吸水鏈霉菌奧薩霉素亞種ATCC15420    QT-00130
長(zhǎng)雙歧桿菌ATCC15707    QT-00131
放射性根瘤菌ATCC15955    QT-00132
短密青霉ATCC16024    QT-00133
副溶血性弧菌ATCC17802    QT-00134
單核增生李斯特菌ATCC19115    QT-00135
缺陷短波單胞菌ATCC19146    QT-00136
師崗鏈霉菌ATCC19166    QT-00137
脫氮付球菌ATCC19367    QT-00138
絲狀鏈霉菌ATCC19753    QT-00139
裂殖壺菌ATCC20888    QT-00140
產(chǎn)朊假絲酵母ATCC22023    QT-00141
乙酸鈣不動(dòng)桿菌ATCC23055    QT-00142
珊瑚鏈霉菌ATCC23901    QT-00143
菲律賓鏈霉菌ATCC23905    QT-00144
里氏木霉ATCC24449    QT-00146
變異鏈球菌ATCC25175    QT-00147
沙鏈霉菌ATCC25488    QT-00148
酪丁酸梭菌ATCC25755    QT-00149
蠣灰鏈霉菌ATCC27455    QT-00150
黃曲霉ATCC28539    QT-00151
噬糖鹽紅菌ATCC29252    QT-00152
隱甲藻ATCC30772    QT-00153
干酪乳桿菌干酪亞種ATCC334    QT-00154
休哈塔假絲酵母ATCC34887    QT-00155
停乳鏈球菌ATCC35666    QT-00156
昆明鏈霉菌ATCC35682    QT-00157
扣囊內(nèi)孢霉ATCC36213    QT-00158
馬克斯克魯維酵母ATCC36534    QT-00159
串珠鐮孢ATCC38946    QT-00160
米曲霉ATCC42149    QT-00161
樹(shù)狀微桿菌ATCC4358    QT-00163
潮濕纖維單胞菌ATCC491    QT-00164
熒光假單胞菌ATCC49642    QT-00165
水螺菌ATCC49643    QT-00166
產(chǎn)氣腸桿菌ATCC49701    QT-00167
串珠鐮孢ATCC52539    QT-00168
金黃色葡萄球菌ATCC6538P    QT-00169
痤瘡丙酸桿菌ATCC6919    QT-00170
鼠李糖乳桿菌ATCC7469    QT-00171
食油假單胞菌ATCC8062    QT-00172
粘質(zhì)沙雷氏菌ATCC8100    QT-00173
蛾微桿菌ATCC8365    QT-00174
糞產(chǎn)堿菌ATCC8750    QT-00175
枯草芽孢桿菌ATCC9327    QT-00176
空腸彎曲桿菌ATCC33291    QT-00177
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Human targeted ribonuclease, TR ELISA Kit
Human protein kinase C,PKC ELISA Kit
Human Aldolase,ALD ELISA Kit
Human L-Lactate Dehydrogenase,L-LDH ELISA Kit
Human acid sphingomyelinase, ASM ELISA Kit
Human arachidonate 5-lipoxygenase,ALOX-5 ELISA Kit
Human 5-lipoxygenase,5-LOLOX ELISA Kit
Human adenylyl cyclase 1,AC-1 ELISA Kit
Human Soluble Adenylate Cyclase,sAC ELISA Kit
Human Ornithine decarboxylase,ODC ELISA Kit
Human protein tyrosine kinase,PTK ELISA Kit
Human protein tyrosine phosphatase,PTPPTPase ELISA Kit
Human TNF α converting enzyme,TACE ELISA Kit
Human Polymorphonuclear Elastase,PMN Elastase ELISA Kit
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Human urokinase-type plasminogen activator,uPAPLAU ELISA Kit
Human Kallikrein 6,KLK 6 ELISA Kit
Human Fructose-1,6-bisphosphate aldolase,FDA ELISA Kit
Human cathepsin D,cath-D ELISA Kit
Human hormone sensitive lipase,HSL ELISA Kit
Human pancreatickininogenase,PK ELISA Kit
Human aconitase 2,ACO-2 ELISA Kit
Human Cyclin-dependent kinase 4,CDK-4 ELISA Kit
Human Placental alkaline pkosphatase,PLAP ELISA Kit
Human receptor tyrosine kinase,RTKs ELISA Kit
Human mitogen activited protein kinase,MAPK ELISA Kit
Human Lysozyme,LZM ELISA Kit
Human focal adhesion kinase,FAK ELISA Kit
Human soluble Phospholipase A2,sPL-A2 ELISA Kit
Human deoxyribonuclease Ⅰ,DNase-Ⅰ ELISA Kit
Human Arginase,Arg ELISA Kit
Human neutrophilic alkaline phosphatase,NAP ELISA Kit
Human anapastic lymphoma kinase,ALK ELISA Kit
Human cytokinin MB,CKMB ELISA Kit
Human myosin light chain kinase,MLCK ELISA Kit
Human glutathione S-transferases,GSTpi ELISA Kit
Human glutathione S-transferases,GSTs ELISA Kit
Human deubiquitinating enzyme,DUB ELISA Kit
Human secreted phospholipase A2,sPLA2 ELISA Kit
Human E3ubiquitin-protein ligase,E3UBPL ELISA Kit
Human mast cell tryptase,MCT ELISA Kit
Human dopachrome tautomerase,DT ELISA Kit
Human E2ubiquitin-conjugating enzyme,E2UBCE ELISA Kit
Human protein disulfide isomerase,PDI ELISA Kit
Human caspase activated deoxyribonuclease,CAD ELISA Kit
Humanγ-glutamyl systeine synthetase,γ-ECS ELISA Kit
Human E1ubiquitin-activating enzyme,E1UBAE ELISA Kit
Human Pepsin,PP ELISA Kit
Human Hexokinase,HK ELISA Kit
Human pyruvate dehydrogenase-E1,PDH E1 ELISA Kit
Human glycogen synthase kinase,GSK ELISA Kit
Human Caspase-9,Casp-9 ELISA Kit
Human Caspase 3,Casp-3 ELISA Kit
Human neutrophil elastase,NE ELISA Kit
Human carbonic anhydrase 2,CA-2 ELISA Kit 
Human A Disintegrin And Metalloprotease 8,ADAM8 ELISA Kit
Human granzymes B,Gzms-B ELISA Kit
Human granzymes A,Gzms-A ELISA Kit
Human Calf intestinal alkaline phosphatase,CIAP ELISA Kit
Human dipeptidyl peptldase Ⅳ,DPPⅣ ELISA Kit
Human Histone Deacetylase,HD ELISA Kit 
Human Methylase ELISA Kit
Human tartrate-resistant acid phosphatase 5b,TRACP-5b ELISA Kit
Human protein kinase B,PKB ELISA Kit
Human heme oxygenase 2,HO-2 ELISA Kit 
Human Pyruvate Kinase,M2-PK ELISA Kit
Human Thyroid-Peroxidase,TPO ELISA Kit
Human alpha-L-fucosidase,AFU ELISA Kit
Human Endothelial nitric oxide synthase,eNOS ELISA Kit
Human heme oxygenase 1,HO-1 ELISA Kit 
Human inducible nitric oxide synthase,iNOS ELISA KIT 
Human omerase,TE ELISA Kit 
Human matrix metalloproteinase 5,MMP-5 ELISA Kit 
Human matrix metalloproteinase 4,MMP-4 ELISA Kit 
Human matrix metalloproteinase 9Gelatinase B,MMP-9 ELISA Kit 
Human Neuron-specific enolase,NSE ELISA Kit
Human Creatine Kinase MB isoenzyme,CK-MB ELISA Kit
Human Prostatic Acid Phosphatase,PAP ELISA Kit
Human Prostaglandin D Synthase,PGDS ELISA Kit
human Pepsinogen C,PG-C ELISA Kit
human Pepsinogen A,PG-A ELISA Kit
Human Urokinase-type Plasminogen Activator Receptor,PLAURuPAR ELISA kit
Human urokinase plasminogen activator,uPA ELISA kit
Human tyrosine kinase with immunoglobulin-like and EGF-like domains 2,Tie-2 ELISA Kit
Human Matrix metalloproteinase 8Neutrophil collagenase,MMP-8 ELISA kit 
Human Matrix metalloproteinase 7,MMP-7 ELISA kit 
Human Matrix metalloproteinase 3,MMP-3 ELISA kit 
Human Matrix metalloproteinase 2Gelatinase A,MMP-2 ELISA kit 
Human Matrix metalloproteinase 13,MMP-13 ELISA kit