"HPC-Y5 epithelioid cells人胰腺導(dǎo)管癌細(xì)胞系 細(xì)胞背景資料:胰腺管癌;淋巴結(jié)轉(zhuǎn)移 細(xì)胞形態(tài):上皮細(xì)胞樣 貼壁細(xì)胞消化傳代時(shí)通常采用兩種方法:一、加入胰酶等細(xì)胞脫落后,再加培養(yǎng)基中止胰酶作用,離心傳代;二、加入胰酶后,鏡下觀察待細(xì)胞始脫落時(shí),棄胰酶,加培養(yǎng)液分瓶。但前者太麻煩,而后者有可能對(duì)細(xì)胞施加胰酶選擇,因?yàn)榭偸琴N壁不牢的細(xì)胞先脫落,對(duì)腫瘤細(xì)胞來說,這部分細(xì)胞有可能是惡性程度較高的細(xì)胞亞群。一種簡(jiǎn)單的消化傳代方法。加入PBS洗去血清或加入胰酶先中和血清的作用(30s),棄之,再加入適量胰酶作用10s-40s(根據(jù)細(xì)胞消化的難易程度),棄之,這樣依賴殘余的胰酶就可將細(xì)胞消化單細(xì)胞。對(duì)于較難消化的細(xì)胞,可以用2%利多卡因消化5-8分鐘,然后再棄去,加培養(yǎng)基吹打也可以,對(duì)細(xì)胞的影響不大。不用PBS也不用Hanks洗,只要把舊培養(yǎng)液吸的干凈一點(diǎn),直接加酶消化應(yīng)該不會(huì)有什么問題。棄培養(yǎng)液后,用0.04%的EDTA沖洗一次,再用1/4v的0.04%的EDTA室溫孵育5min,棄取大部分EDTA,加入與剩余EDTA等量的胰酶(預(yù)熱)總體積1/10v。消化到有細(xì)胞脫落。不過有人說EDTA對(duì)細(xì)胞不好,有證據(jù)嗎?培養(yǎng)的BASMC:倒掉舊培養(yǎng)液加入少量胰酶沖一下,倒掉再加入0.125-0.25%胰酶約6-10滴或1ml(25ml bottle)消化再加入適量新培養(yǎng)基中和,并分瓶這種方法簡(jiǎn)單、省事;效果很好并且不損失細(xì)胞! HRMC細(xì)胞系相關(guān)產(chǎn)品:Institute for Medical Research-90細(xì)胞、3T3-Swiss細(xì)胞、Hs852T細(xì)胞 H2330細(xì)胞系相關(guān)產(chǎn)品:MeT-5A細(xì)胞、OEC19細(xì)胞、hTERTHME1細(xì)胞 MD Anderson-Metastatic Breast-157細(xì)胞系相關(guān)產(chǎn)品:MDCKII細(xì)胞、NCI-HUT-596細(xì)胞、SK-ES細(xì)胞 細(xì)胞生長(zhǎng):貼壁 HPC-Y5 epithelioid cells人胰腺導(dǎo)管癌細(xì)胞系 細(xì)胞物種來源:人源或鼠源等其它物種來源 細(xì)胞產(chǎn)品包裝:復(fù)蘇形式:T25培養(yǎng)瓶(一瓶)或凍存形式:1ml凍存管(兩支) 冷凍細(xì)胞解凍程序:依據(jù)細(xì)胞株數(shù)據(jù)單指定之基礎(chǔ)培養(yǎng)基種類、血清種類和其它指定之成份和比例,制備培養(yǎng)基。絕大多數(shù)之細(xì)胞均無法立即適應(yīng)不同之基礎(chǔ)培養(yǎng)基或不同之血清種類,若因?qū)嶒?yàn)需要,必須有所不同時(shí),務(wù)必以緩慢比例漸次改變培養(yǎng)基組成,確定細(xì)胞適應(yīng)后,方進(jìn)行所需之實(shí)驗(yàn)。FBS(fetal bovine serum,胚牛血清), CS(calf serum,小牛血清)和HS(horse serum,馬血清),對(duì)細(xì)胞而言差異極大,請(qǐng)務(wù)必依據(jù)細(xì)胞株資料單指定之血清種類培養(yǎng)之。將培養(yǎng)基置于37°C水槽中回溫,回溫后噴以70%酒精并擦拭之,移入無菌操作臺(tái)內(nèi)。取出冷凍管,立即放入37°C水槽中快速解凍,水面高度不可接近或高過冷凍管之蓋沿,否則易發(fā)生污染。輕搖冷凍管使其在1分鐘內(nèi)全部融化后,以70%ethanol擦拭冷凍管外部,移入無菌操作臺(tái)內(nèi)。依據(jù)細(xì)胞種類和濃度,于無菌操作臺(tái)內(nèi)?。保癿l培養(yǎng)基加至T25或T75flask中。取出已解凍之細(xì)胞懸浮液,緩緩加入T25或T75flask內(nèi)之培養(yǎng)基,混合均勻,放入37°C,5%CO2培養(yǎng)箱培養(yǎng)。對(duì)絕大多數(shù)細(xì)胞而言,1%以下之冷凍保護(hù)劑DMSO,不會(huì)對(duì)細(xì)胞之貼附或活化有不良影響,不需立刻由解凍細(xì)胞中去除,待第二天確定細(xì)胞生長(zhǎng)或貼附良好后再去除即可。惟對(duì)極少數(shù)因?qū)MSO敏感或會(huì)造成細(xì)胞分化之細(xì)胞,需立即去除DMSO者,則可將解凍后之細(xì)胞懸浮液放入5-10ml培養(yǎng)基中,離心300xg(約1000rpm),5分鐘,小心移去上清液,加入適量新鮮培養(yǎng)基,將細(xì)胞均勻混合后,轉(zhuǎn)移至培養(yǎng)瓶中,再放入37°C,5%CO2培養(yǎng)箱培養(yǎng)。 細(xì)胞傳代方法:1:2-1:3傳代;每周換液2-3次。 WILL-2細(xì)胞系相關(guān)產(chǎn)品:Baby Hamster Kidney from litter No. 21細(xì)胞、BMF細(xì)胞、NCIH2172細(xì)胞 SU-DHL-8細(xì)胞系相關(guān)產(chǎn)品:CII細(xì)胞、HMrSV5細(xì)胞、MCA38細(xì)胞 Hep 3B2.1-7細(xì)胞系相關(guān)產(chǎn)品:MDAMB134細(xì)胞、LAPC4細(xì)胞、BT-325細(xì)胞 T/GHA-VSMC細(xì)胞系相關(guān)產(chǎn)品:SGC-7901細(xì)胞、NCIH441細(xì)胞、KOSC2細(xì)胞 P560細(xì)胞系相關(guān)產(chǎn)品:HuT-78細(xì)胞、H889細(xì)胞、OVCAR-10細(xì)胞 細(xì)胞來源說明:細(xì)胞主要來源ATCC、DSMZ、ECACC、RIKEN、INCell、promocell、ScienCell、ECACC、JCRB、KCLB、Asterand、ICLC以及少數(shù)國外著名大學(xué)建系 HPC-Y5 epithelioid cells人胰腺導(dǎo)管癌細(xì)胞系 細(xì)胞生長(zhǎng)特性:貼壁 細(xì)胞形態(tài)特性:詳見細(xì)胞說明書 細(xì)胞常規(guī)培養(yǎng)傳代流程(請(qǐng)嚴(yán)格遵照無菌操作):1)吸出原培養(yǎng)瓶中的培養(yǎng)基,PBS緩沖液潤(rùn)洗細(xì)胞兩次,加2-3ml 0.25%胰酶進(jìn)行消化細(xì)胞(注意把握消化時(shí)間,通??刂圃冢?2min);2)鏡下觀察消化情況,在細(xì)胞邊緣縮小,貼壁松動(dòng)時(shí)(不建議消化到細(xì)胞漂浮)去掉胰酶,加6-8ml完全培養(yǎng)基,輕輕吹打細(xì)胞層,盡量把細(xì)胞層吹落,吹散;3)取部分細(xì)胞懸液轉(zhuǎn)移到新的培養(yǎng)皿/瓶中,添加適當(dāng)?shù)耐耆囵B(yǎng)基,把細(xì)胞懸液打勻,于培養(yǎng)箱中培養(yǎng);4)注意培養(yǎng)基PH值變化情況,定期換液(每周2-3次),待細(xì)胞密度達(dá)到80%以后重復(fù)1項(xiàng)操作或者凍存;特別注意:(如使用公共實(shí)驗(yàn)室或者初次接觸細(xì)胞培養(yǎng),建議添加雙抗培養(yǎng))1)收到細(xì)胞后請(qǐng)盡快更換為含15%血清的新鮮培養(yǎng)基,如因特殊情況需要繼續(xù)使用原瓶,請(qǐng)?jiān)谠颗囵B(yǎng)基中額外添加10%的血清,(原瓶培養(yǎng)基的繼續(xù)使用時(shí)間最長(zhǎng)不宜超過72小時(shí));2)貼壁細(xì)胞收到當(dāng)天切忌立刻消化,請(qǐng)將細(xì)胞換液后放置培養(yǎng)箱孵育到第二天再做消化傳代,請(qǐng)優(yōu)先選擇直徑6cm的培養(yǎng)皿進(jìn)行傳代培養(yǎng);3)如簽收時(shí)出現(xiàn)培養(yǎng)瓶壁破裂,漏液等情況請(qǐng)及時(shí)做好照片記錄并聯(lián)系實(shí)驗(yàn)室。 RPMI-2650細(xì)胞系相關(guān)產(chǎn)品:BC-021細(xì)胞、NHLF細(xì)胞、H-1648細(xì)胞 OC 316細(xì)胞系相關(guān)產(chǎn)品:SF-539 BT細(xì)胞、Nittby-Salford 1細(xì)胞、MNNG-HOS (Cl#5)細(xì)胞 B6Tert-1細(xì)胞系相關(guān)產(chǎn)品:CEM-0細(xì)胞、EL4細(xì)胞、SNT8細(xì)胞 Hepa1-6細(xì)胞系相關(guān)產(chǎn)品:Dx5細(xì)胞、CATH-a細(xì)胞、SAS細(xì)胞 VK2細(xì)胞系相關(guān)產(chǎn)品:COR-L26細(xì)胞、H2170細(xì)胞、H2171細(xì)胞 NCI-H526細(xì)胞系相關(guān)產(chǎn)品:P3-X63-Ag8.653細(xì)胞、HS-940-T細(xì)胞、MCMEC細(xì)胞 MN9D細(xì)胞系相關(guān)產(chǎn)品:U251n細(xì)胞、Vero from pool #76細(xì)胞、HAPI細(xì)胞 H-2227細(xì)胞系相關(guān)產(chǎn)品:HT1197細(xì)胞、HUT 125細(xì)胞、J.E6-1細(xì)胞 HCC-202細(xì)胞系相關(guān)產(chǎn)品:RT4細(xì)胞、NB1-RGB細(xì)胞、ND7/23細(xì)胞 IPEC-J2細(xì)胞系相關(guān)產(chǎn)品:NT2細(xì)胞、Det. 562細(xì)胞、ACC-M細(xì)胞 HPC-Y5 epithelioid cells人胰腺導(dǎo)管癌細(xì)胞系 KYSE70細(xì)胞系相關(guān)產(chǎn)品:TF-1細(xì)胞、H-1838細(xì)胞、52PE細(xì)胞 HCC1937細(xì)胞系相關(guān)產(chǎn)品:JROECL 21細(xì)胞、SHSY-5Y細(xì)胞、DMS53細(xì)胞 CHOK1細(xì)胞系相關(guān)產(chǎn)品:SKRC20細(xì)胞、R 1610細(xì)胞、KHYG-1細(xì)胞 Calf Pulmonary Artery Endothelial細(xì)胞系相關(guān)產(chǎn)品:GTL16細(xì)胞、T47-D細(xì)胞、MDA-468細(xì)胞 PA I細(xì)胞系相關(guān)產(chǎn)品:GM00637H細(xì)胞、P3J.HR1K細(xì)胞、EU-2細(xì)胞 HEK-293-EBNA細(xì)胞系相關(guān)產(chǎn)品:32Dcl3細(xì)胞、Bac 1.2F5細(xì)胞、BALB/3T3 clone A31細(xì)胞 HEK-293-EBNA細(xì)胞系相關(guān)產(chǎn)品:32Dcl3細(xì)胞、Bac 1.2F5細(xì)胞、BALB/3T3 clone A31細(xì)胞 F442-A細(xì)胞系相關(guān)產(chǎn)品:MOLT-16細(xì)胞、DH 82細(xì)胞、UCLA-SO-M14細(xì)胞 YH細(xì)胞系相關(guān)產(chǎn)品:NL-20細(xì)胞、Virginia Mason Research Center-Lung Cancer D細(xì)胞、NCIH596細(xì)胞 MC-38細(xì)胞系相關(guān)產(chǎn)品:OE-21細(xì)胞、SUDHL8細(xì)胞、HeLa S3細(xì)胞 SNU387細(xì)胞系相關(guān)產(chǎn)品:3T3L1細(xì)胞、SNU739細(xì)胞、Human Intestinal Epithelial Cell-6細(xì)胞 COLO 205細(xì)胞系相關(guān)產(chǎn)品:H4-II-E細(xì)胞、SUM 102PT細(xì)胞、COLO_320DM細(xì)胞 BA/F3細(xì)胞系相關(guān)產(chǎn)品:DSL-6A-C1細(xì)胞、HEP-3B2細(xì)胞、MDA-MB468細(xì)胞 N1E115細(xì)胞系相關(guān)產(chǎn)品:ST-2細(xì)胞、Calu-6細(xì)胞、Kit225 K6細(xì)胞 H-28細(xì)胞系相關(guān)產(chǎn)品:NCIH2126細(xì)胞、NCI-H2228細(xì)胞、UACC-812細(xì)胞 RGCs細(xì)胞系相關(guān)產(chǎn)品:SK-MES-1細(xì)胞、C81-61細(xì)胞、3T6 Swiss Albino細(xì)胞 H2452細(xì)胞系相關(guān)產(chǎn)品:BrCL12細(xì)胞、PTK2細(xì)胞、EB2細(xì)胞 KYSE30細(xì)胞系相關(guān)產(chǎn)品:MCF7/WT細(xì)胞、Lymph Node Carcinoma of the prostate細(xì)胞、HCA7細(xì)胞 SUD-4細(xì)胞系相關(guān)產(chǎn)品:CHOK1細(xì)胞、H4-II-E-C3細(xì)胞、CHL-IU細(xì)胞 H441細(xì)胞系相關(guān)產(chǎn)品:HHL5細(xì)胞、H748細(xì)胞、MNNG/HOS Clone F-5細(xì)胞 STTG1細(xì)胞系相關(guān)產(chǎn)品:QGY 7701細(xì)胞、LNCaP-Clone-FGC細(xì)胞、SKRC20細(xì)胞 PLA-801D細(xì)胞系相關(guān)產(chǎn)品:THP1細(xì)胞、PMVEC細(xì)胞、NCIH64細(xì)胞 Hs840T細(xì)胞系相關(guān)產(chǎn)品:BTI-Tn5B14細(xì)胞、NCI H548細(xì)胞、HT-55細(xì)胞 EVSAT細(xì)胞系相關(guān)產(chǎn)品:MB49細(xì)胞、HCC0044細(xì)胞、PVEC細(xì)胞 DMS 273細(xì)胞系相關(guān)產(chǎn)品:AN-3細(xì)胞、OCI-Ly7細(xì)胞、Hs940-T細(xì)胞 U-118MG細(xì)胞系相關(guān)產(chǎn)品:PK-15細(xì)胞、293EBNA細(xì)胞、HPDEC細(xì)胞 TGBC11T細(xì)胞系相關(guān)產(chǎn)品:Co 205細(xì)胞、CLONE M3細(xì)胞、CMT167細(xì)胞 UCLA SO M21細(xì)胞系相關(guān)產(chǎn)品:MAVER-1細(xì)胞、Mono Mac 6細(xì)胞、SF-295細(xì)胞 CMT-64細(xì)胞系相關(guān)產(chǎn)品:GP2d細(xì)胞、tdott細(xì)胞、UMNSAH-DF1細(xì)胞 Panc02.03細(xì)胞系相關(guān)產(chǎn)品:H2198細(xì)胞、U118MG細(xì)胞、UWB1-289細(xì)胞 EoL-1-cell細(xì)胞系相關(guān)產(chǎn)品:BRL細(xì)胞、H-1048細(xì)胞、SPCA-1細(xì)胞 SaOS細(xì)胞系相關(guān)產(chǎn)品:EA.Hy926細(xì)胞、RPMI #8226細(xì)胞、CWR22R-V1細(xì)胞 NTERA-2 cl.D1細(xì)胞系相關(guān)產(chǎn)品:GM02219細(xì)胞、HMC3細(xì)胞、B16/F1細(xì)胞 JROECL 33細(xì)胞系相關(guān)產(chǎn)品:HOCF細(xì)胞、BHK細(xì)胞、C3H10T1/2CL8細(xì)胞 ECC1細(xì)胞系相關(guān)產(chǎn)品:MC3T3-E1 Subclone 24細(xì)胞、GM-3573細(xì)胞、DKMG細(xì)胞 Hut292細(xì)胞系相關(guān)產(chǎn)品:H650細(xì)胞、GM3569細(xì)胞、H-841細(xì)胞 OK細(xì)胞系相關(guān)產(chǎn)品:Y3-Ag123細(xì)胞、C-8161細(xì)胞、KU 812細(xì)胞 NALM-6-M1細(xì)胞系相關(guān)產(chǎn)品:HCC-15細(xì)胞、KRCY細(xì)胞、LAD 2細(xì)胞 NALM-6-M1細(xì)胞系相關(guān)產(chǎn)品:HCC-15細(xì)胞、KRCY細(xì)胞、LAD 2細(xì)胞 MDA-453細(xì)胞系相關(guān)產(chǎn)品:JCA1細(xì)胞、CHP-100細(xì)胞、Caco-2/BBe 1細(xì)胞 Tokyo Medical and Dental university 8細(xì)胞系相關(guān)產(chǎn)品:CEK細(xì)胞、HB 611細(xì)胞、FLC-7細(xì)胞 NCI-H1623細(xì)胞系相關(guān)產(chǎn)品:COLO 320HSR細(xì)胞、HHUA細(xì)胞、ST486細(xì)胞 SNU-761細(xì)胞系相關(guān)產(chǎn)品:HM1900細(xì)胞、MES 13細(xì)胞、769P細(xì)胞 WEHI3B細(xì)胞系相關(guān)產(chǎn)品:OS-732細(xì)胞、HepG2/C3A細(xì)胞、Madison細(xì)胞 PG [Human lung carcinoma]細(xì)胞系相關(guān)產(chǎn)品:NCIH2009細(xì)胞、LWnt3A細(xì)胞、CL11細(xì)胞 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