"Farage Lymphoblastoid cells人B淋巴瘤細(xì)胞系 細(xì)胞背景資料:Farage細(xì)胞源于一名患有彌漫性大B細(xì)胞淋巴瘤(DLBCL)白人女性的活檢淋巴組織,由HBen-Bassat建系。經(jīng)IL-4處理,該細(xì)胞CD21,CD22,CD54和CD58表達(dá)上調(diào),而CD21,CD22,andCD38表達(dá)下調(diào)。 細(xì)胞形態(tài):淋巴母細(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(lái)說(shuō),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(wèn)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(guò)有人說(shuō)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! HCMEC(BL12-H)細(xì)胞系相關(guān)產(chǎn)品:OVCAR10細(xì)胞、NCI-H1404細(xì)胞、AMO-1細(xì)胞 95D細(xì)胞系相關(guān)產(chǎn)品:OCUM-1細(xì)胞、C4-2細(xì)胞、NTHY-ORI3.1細(xì)胞 NCM356細(xì)胞系相關(guān)產(chǎn)品:MDCK II細(xì)胞、REC1細(xì)胞、AK細(xì)胞 細(xì)胞生長(zhǎng):懸浮 Farage Lymphoblastoid cells人B淋巴瘤細(xì)胞系 細(xì)胞物種來(lái)源:人源或鼠源等其它物種來(lái)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) 血清必須貯存于–20~-70℃,若存放于4℃,請(qǐng)勿超過(guò)一個(gè)月。如果一次無(wú)法用完一瓶,可將40~45ml分裝于無(wú)菌50ml離心管中,由于血清結(jié)凍時(shí)體積會(huì)增加約10%,必須預(yù)留此膨脹體積之空間,否則易發(fā)生污染或容器凍裂之情形。一般廠商提供之血清為無(wú)菌,不需再無(wú)菌過(guò)濾。若發(fā)現(xiàn)血清有許多懸浮物,則可將血清加入培養(yǎng)基內(nèi)一起過(guò)濾,勿直接過(guò)濾血清。瓶裝(500ml)血清解凍步驟(逐步解凍法):-20℃或–70℃至4℃冰箱溶解一天,至室溫下全溶后再分裝,一般以50ml無(wú)菌離心管可分裝40~45ml。在溶解過(guò)程中須規(guī)則搖晃均勻(小心勿造成氣泡),使溫度與成分均一,減少沈淀的發(fā)生。勿直接由–20℃直接至37℃解凍,因溫度改變太大,容易造成蛋白質(zhì)凝結(jié)而發(fā)生沈淀。勿將血清置于37℃太久,若在37℃放置太久,血清會(huì)變得混濁,同時(shí)血清中許多較不穩(wěn)定之成份亦會(huì)因此受到破壞,而影響血清之品質(zhì)。血清凝絮物:發(fā)生之原因有許多種,但普遍之原因是血清中之脂蛋白(lipoprotein) 變性及解凍后血清中存在之血纖維蛋白(fibrin) 造成,這些凝絮沈淀物不會(huì)影響血清本身之品質(zhì)。若欲減少這些凝絮沈淀物,可用離心3000rpm,5min去除,或離心后上清液可以加入培養(yǎng)基中一起過(guò)濾。不建議用過(guò)濾步驟去除這些凝絮沈淀物,因?yàn)闀?huì)阻塞過(guò)濾膜。顯微鏡下觀察之“小黑點(diǎn)”:通常經(jīng)過(guò)熱處理之血清,沈淀物的形成會(huì)顯著的增多。有些沈淀物在顯微鏡下觀察像是“小黑點(diǎn)”,常會(huì)誤認(rèn)為血清遭受污染,而將血清放在37℃中欲培養(yǎng)此“微生物”,但在37℃環(huán)境下,又會(huì)使此沈淀物增多,更會(huì)誤認(rèn)為微生物之增殖,但以培養(yǎng)細(xì)菌之培養(yǎng)基檢測(cè),又沒(méi)有污染。一般而言,此小黑點(diǎn)應(yīng)不會(huì)影響細(xì)胞之生長(zhǎng),但若懷疑此血清之品質(zhì),應(yīng)立即停用,更換另一批號(hào)的血清。 細(xì)胞傳代方法:1:3傳代,2-3天傳一代 EOL1細(xì)胞系相關(guān)產(chǎn)品:H-2126細(xì)胞、NCI-SNU-475細(xì)胞、CMT.64細(xì)胞 Hs-274-T細(xì)胞系相關(guān)產(chǎn)品:GI-1細(xì)胞、HeLa S-3細(xì)胞、Pfeiffer細(xì)胞 A-375SM細(xì)胞系相關(guān)產(chǎn)品:HCa/16A3-F細(xì)胞、NCIH1355細(xì)胞、NR8383細(xì)胞 H1666_DA細(xì)胞系相關(guān)產(chǎn)品:RK13細(xì)胞、NCI-SNU-387細(xì)胞、ARO細(xì)胞 M-NFS-60細(xì)胞系相關(guān)產(chǎn)品:ACC-2細(xì)胞、EOL1細(xì)胞、Caco-2 BBe細(xì)胞 細(xì)胞來(lái)源說(shuō)明:細(xì)胞主要來(lái)源ATCC、DSMZ、ECACC、RIKEN、INCell、promocell、ScienCell、ECACC、JCRB、KCLB、Asterand、ICLC以及少數(shù)國(guó)外著名大學(xué)建系 Farage Lymphoblastoid cells人B淋巴瘤細(xì)胞系 細(xì)胞生長(zhǎng)特性:懸浮生長(zhǎng) 細(xì)胞形態(tài)特性:淋巴母細(xì)胞樣 細(xì)胞常規(guī)培養(yǎng)傳代流程(請(qǐng)嚴(yán)格遵照無(wú)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(guò)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。 DMS-114細(xì)胞系相關(guān)產(chǎn)品:MDAMB-231細(xì)胞、253J細(xì)胞、293AD細(xì)胞 H1975細(xì)胞系相關(guān)產(chǎn)品:GM0637細(xì)胞、Spodoptera frugiperda clone 9細(xì)胞、Human Epidermoid carcinoma #2細(xì)胞 Baby Hamster Kidney from litter No. 21細(xì)胞系相關(guān)產(chǎn)品:PCI-4B細(xì)胞、KATO III細(xì)胞、GFP-Olig2細(xì)胞 HCC-1359細(xì)胞系相關(guān)產(chǎn)品:SUM-159PT細(xì)胞、NIH:OVCAR-4細(xì)胞、Karpas-299細(xì)胞 BxPC-3細(xì)胞系相關(guān)產(chǎn)品:RPMI-7666細(xì)胞、M 1細(xì)胞、H-345細(xì)胞 P19細(xì)胞系相關(guān)產(chǎn)品:KYSE140細(xì)胞、LN-18細(xì)胞、L 363細(xì)胞 Ontario Cancer Institute-Acute Myeloid Leukemia-3細(xì)胞系相關(guān)產(chǎn)品:SNU-869細(xì)胞、SK-Col-1細(xì)胞、D341細(xì)胞 WTRL1細(xì)胞系相關(guān)產(chǎn)品:AAV-293細(xì)胞、H-920細(xì)胞、DanG細(xì)胞 52PE細(xì)胞系相關(guān)產(chǎn)品:Panc04.03細(xì)胞、BT-20細(xì)胞、Instituto Biologico-Rim Suino-2細(xì)胞 SKG IIIa細(xì)胞系相關(guān)產(chǎn)品:HS578T細(xì)胞、NIH:OVCAR-3細(xì)胞、WEHI 3細(xì)胞 Farage Lymphoblastoid cells人B淋巴瘤細(xì)胞系 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 GT1-1細(xì)胞系相關(guān)產(chǎn)品:LY-R細(xì)胞、BCaP 37細(xì)胞、2B4細(xì)胞 RSC-364細(xì)胞系相關(guān)產(chǎn)品:NCI-H-128細(xì)胞、KMBC細(xì)胞、MES-SA細(xì)胞 HEP-3B2細(xì)胞系相關(guān)產(chǎn)品:CNE-2Z細(xì)胞、OCLY8細(xì)胞、KMM-1細(xì)胞 PL9細(xì)胞系相關(guān)產(chǎn)品:Centre Antoine Lacassagne-78細(xì)胞、CAKI1細(xì)胞、MDA-MB-231-GFP細(xì)胞 NSC34細(xì)胞系相關(guān)產(chǎn)品:NTera 2/cl.D1細(xì)胞、H.Ep. #2細(xì)胞、LC1-Sq細(xì)胞 16HBE細(xì)胞系相關(guān)產(chǎn)品:MC38細(xì)胞、RASMC細(xì)胞、SNB.19細(xì)胞 Ramos-2G6-4C10細(xì)胞系相關(guān)產(chǎn)品:BT 549細(xì)胞、PANC0327細(xì)胞、COS7細(xì)胞 PAN 02細(xì)胞系相關(guān)產(chǎn)品:MFM-223細(xì)胞、MTEC1細(xì)胞、RAW-264.7細(xì)胞 MIN6細(xì)胞系相關(guān)產(chǎn)品:NCIH1568細(xì)胞、10RGB細(xì)胞、LS 123細(xì)胞 Lu-65細(xì)胞系相關(guān)產(chǎn)品:LO2細(xì)胞、NCIH661細(xì)胞、MDAMB175細(xì)胞 Pro-5 Lec1.3c細(xì)胞系相關(guān)產(chǎn)品:HuH28細(xì)胞、VM Cub 1細(xì)胞、HS-294T細(xì)胞 H2135細(xì)胞系相關(guān)產(chǎn)品:HCC-1143細(xì)胞、Pa14C細(xì)胞、OCI-Ly19細(xì)胞 SW1222細(xì)胞系相關(guān)產(chǎn)品:HRA 19細(xì)胞、Normal Rat Kidney-49F細(xì)胞、HCC941122細(xì)胞 CATH-a細(xì)胞系相關(guān)產(chǎn)品:M210B4細(xì)胞、HCFB(HCF)細(xì)胞、Hs-688A-T細(xì)胞 TE-6細(xì)胞系相關(guān)產(chǎn)品:SK MEL 28細(xì)胞、MFC細(xì)胞、REC-1細(xì)胞 NKT細(xì)胞系相關(guān)產(chǎn)品:MASMC細(xì)胞、Tca8113細(xì)胞、SKMEL5細(xì)胞 Bovine Turbinate細(xì)胞系相關(guān)產(chǎn)品:MARC-145細(xì)胞、TCC-PAN2細(xì)胞、RPMI-1846細(xì)胞 251 MG細(xì)胞系相關(guān)產(chǎn)品:2BS細(xì)胞、NCIH1618細(xì)胞、Hep G2/C3A細(xì)胞 Hs-578Bst細(xì)胞系相關(guān)產(chǎn)品:MN 60細(xì)胞、P3-Jiyoye細(xì)胞、UMR-106細(xì)胞 D283細(xì)胞系相關(guān)產(chǎn)品:SKMEL3細(xì)胞、Intestinal Epithelioid Cell line No. 18細(xì)胞、Det 562細(xì)胞 HeLa/S3細(xì)胞系相關(guān)產(chǎn)品:Lilly Laboratories Cell-Porcine Kidney 1細(xì)胞、NCTC1469細(xì)胞、GP-293細(xì)胞 Hs-343-T細(xì)胞系相關(guān)產(chǎn)品:FL83B細(xì)胞、RCK-8細(xì)胞、HCT FET細(xì)胞 RPMI 8226/S細(xì)胞系相關(guān)產(chǎn)品:AC16 [Human hybrid]細(xì)胞、NCIH1650細(xì)胞、PANC-1細(xì)胞 TE1細(xì)胞系相關(guān)產(chǎn)品:H-2228細(xì)胞、COLO-320HSR細(xì)胞、Med 341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 Melan-a細(xì)胞系相關(guān)產(chǎn)品:HCC15細(xì)胞、HNE1細(xì)胞、HPAF-2細(xì)胞 RWPE-2細(xì)胞系相關(guān)產(chǎn)品:C 643細(xì)胞、SF-539細(xì)胞、NT2D1細(xì)胞 COR-L88細(xì)胞系相關(guān)產(chǎn)品:FLC-7細(xì)胞、JCA1細(xì)胞、H-1385細(xì)胞 BJA-B細(xì)胞系相關(guān)產(chǎn)品:Y3Ag1.2.3細(xì)胞、NE-1細(xì)胞、BC-020細(xì)胞 OV1063細(xì)胞系相關(guān)產(chǎn)品:KYSE-30細(xì)胞、Colon38細(xì)胞、KMS-18細(xì)胞 KP-N-RT-BM-1細(xì)胞系相關(guān)產(chǎn)品:OCIAML3細(xì)胞、KAT5細(xì)胞、578T細(xì)胞 SU 86.86細(xì)胞系相關(guān)產(chǎn)品:UWB1.289細(xì)胞、HEK-293 c18細(xì)胞、Hs840_T細(xì)胞 SW-1353細(xì)胞系相關(guān)產(chǎn)品:AZ521細(xì)胞、NCIH146細(xì)胞、C42細(xì)胞 Vero 76 clone E-6細(xì)胞系相關(guān)產(chǎn)品:SK HEP 01細(xì)胞、JEKO細(xì)胞、SKNSH細(xì)胞 CHO/dhFr-細(xì)胞系相關(guān)產(chǎn)品:MDBK細(xì)胞、NOK細(xì)胞、HT 115細(xì)胞 Hs-746T細(xì)胞系相關(guān)產(chǎn)品:NOZAWA細(xì)胞、CCRF/CEM-C7細(xì)胞、EAhy926細(xì)胞 hSCC-25細(xì)胞系相關(guān)產(chǎn)品:SF 295細(xì)胞、K1735細(xì)胞、HDF-a細(xì)胞 MV4;11細(xì)胞系相關(guān)產(chǎn)品:Transformed Human Liver Epithelial-2細(xì)胞、LICR-HN6細(xì)胞、PA1細(xì)胞 DI-TNC1細(xì)胞系相關(guān)產(chǎn)品:SN12CPM6細(xì)胞、Baby Hamster Kidney from litter No. 21細(xì)胞、hEM15A細(xì)胞 KU.812細(xì)胞系相關(guān)產(chǎn)品:J-774A.1細(xì)胞、Hi-five細(xì)胞、HEK-293-H細(xì)胞 H-1819細(xì)胞系相關(guān)產(chǎn)品:5637細(xì)胞、SKNDZ細(xì)胞、OCI-Ly3細(xì)胞 NIH:OVCAR-10細(xì)胞系相關(guān)產(chǎn)品:RPTEC TERT1細(xì)胞、HONE-1細(xì)胞、MDA MB 175 VII細(xì)胞 HHFK細(xì)胞系相關(guān)產(chǎn)品:HEMCSS細(xì)胞、MDA-MB-134細(xì)胞、Michigan Cancer Foundation-10A細(xì)胞 D-324 Med細(xì)胞系相關(guān)產(chǎn)品:NCI-SNU-182細(xì)胞、HCT-116細(xì)胞、H2066細(xì)胞 NCIH1048細(xì)胞系相關(guān)產(chǎn)品:SW-1573細(xì)胞、OCIAML5細(xì)胞、MDCC-MSB1細(xì)胞 H719細(xì)胞系相關(guān)產(chǎn)品:CAL-78細(xì)胞、MV411細(xì)胞、HuT 78細(xì)胞 HAPI細(xì)胞系相關(guān)產(chǎn)品:ARO細(xì)胞、H1944細(xì)胞、NCIH2227細(xì)胞 Hs-729-T細(xì)胞系相關(guān)產(chǎn)品:RPMI8226細(xì)胞、OCILY-10細(xì)胞、MM96L細(xì)胞 159PT細(xì)胞系相關(guān)產(chǎn)品:NCI-H2122細(xì)胞、SuDHL 8細(xì)胞、CCD-1112sk細(xì)胞 32D CL3細(xì)胞系相關(guān)產(chǎn)品:NCI-H87細(xì)胞、Colo-201細(xì)胞、DAKIKI Clone 1細(xì)胞 Farage Lymphoblastoid cells人B淋巴瘤細(xì)胞系 10RGB細(xì)胞系相關(guān)產(chǎn)品:OCM1A細(xì)胞、TKB-1細(xì)胞、CEM-T4細(xì)胞 SKG IIIa細(xì)胞系相關(guān)產(chǎn)品:HS578T細(xì)胞、HSC4細(xì)胞、KP-N-RT-BM-1細(xì)胞 T47D:A細(xì)胞系相關(guān)產(chǎn)品:PL-45細(xì)胞、C17細(xì)胞、PCI:SG231細(xì)胞 Baby Hamster Kidney from litter No. 21細(xì)胞系相關(guān)產(chǎn)品:PCI-4B細(xì)胞、H3396細(xì)胞、HOP-62細(xì)胞 "
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