"HBL-1 [Human diffuse large B-cell lymphoma] Lymphoblastoid cells人彌漫大B淋巴瘤細(xì)胞系
細(xì)胞背景資料:彌漫大B淋巴瘤;男性
細(xì)胞形態(tài):淋巴母細(xì)胞樣
細(xì)胞凍存復(fù)蘇材料與方法步驟:常用的細(xì)胞冷凍貯存器為液氮貯存器,規(guī)格有35L和50L兩種。使用時(shí)要注意以下幾點(diǎn):(1)一般兩周需充液氮一次,至少一個(gè)月充氮一次。液氮溫度達(dá)-196℃,使用時(shí)注意勿讓液氮濺到皮膚上,以免引起凍傷。(2)液氮容器為雙層結(jié)構(gòu),中間為真空層,瓶口有雙層焊接處,應(yīng)防止焊接部裂開(kāi)。(3)在裝入液氮時(shí),要注意緩慢小心,并用厚紙卷筒或特制漏斗作引導(dǎo),使液氮直達(dá)瓶底,如有專用液氮灌注裝置則更好。若為初次使用,加液氮時(shí)更要緩慢,以免溫度驟降而使容器損壞。細(xì)胞凍存時(shí)常備的材料有:0.25%胰蛋白酶,含10%~20%的血清培養(yǎng)液,DMSO(分析純)或無(wú)色新鮮甘油(121°C蒸氣高壓消毒),2mL安瓿(或?qū)S眉?xì)胞凍存管)、吸管、離心管、噴燈、紗布袋(或凍存管架)等。主要操作步驟為:(1)選擇處于對(duì)數(shù)生長(zhǎng)期的細(xì)胞,在凍存前一天換液。將多個(gè)培養(yǎng)瓶中的細(xì)胞培養(yǎng) 液去掉,用0.25%胰蛋白酶消化。適時(shí)去掉胰蛋白酶,加入少量新培養(yǎng)液。用吸管吸取培養(yǎng)液反復(fù)吹打瓶壁上的細(xì)胞,使其成為均勻分散的細(xì)胞懸液。懸浮生產(chǎn)細(xì)胞則不要消化處理。然后將細(xì)胞收集于離心管中離心(1000r/min,10分鐘)。(2)去上清液,加入含20%小牛血清的完全培養(yǎng)基,于4℃預(yù)冷15分鐘后,逐滴加入已無(wú)菌的DMSO或甘油,用吸管輕輕吹打使細(xì)胞均勻,細(xì)胞濃度為3×106~1×107/mL之間。(3)將上述細(xì)胞分裝于安瓿或?qū)S美鋬鏊芰瞎苤?,安瓿裝1~1.5mL在火焰噴燈上封口,封口處要完全封閉,圓滑無(wú)勾。冷凍管要將蓋子蓋緊,并標(biāo)記好細(xì)胞名稱和凍存日期,同時(shí)作好登記(日期、細(xì)胞種類及代次、凍存支數(shù))。(4)將裝好細(xì)胞的安瓿或凍存管裝入沙布袋內(nèi);置于液氮容器頸口處存放過(guò)夜,次日轉(zhuǎn)入液氮中。采用控制降溫速度的方法也可采用下列步驟:先將安瓿置入4℃冰箱中2~3小時(shí),再移至冰箱冷凍室內(nèi)3~4小時(shí),再吊入液氮容器頸氣態(tài)部分存放2小時(shí),最后沉入液氮中。細(xì)胞凍存在液氮中可以長(zhǎng)期保存,但為妥善起見(jiàn),凍存半年后,取出一只安瓿細(xì)胞復(fù)蘇培養(yǎng),觀察生長(zhǎng)情況,然后再繼續(xù)凍存。
P3-X63.Ag8.653細(xì)胞系相關(guān)產(chǎn)品:MonoMac 6細(xì)胞、SU-DHL-6細(xì)胞、BEL7405細(xì)胞
CV-1.K細(xì)胞系相關(guān)產(chǎn)品:OVMANA細(xì)胞、Ku812F細(xì)胞、RH8994細(xì)胞
H1395細(xì)胞系相關(guān)產(chǎn)品:MIO-M1細(xì)胞、Hs840_T細(xì)胞、Caco2BBe細(xì)胞
細(xì)胞生長(zhǎng):懸浮
HBL-1 [Human diffuse large B-cell lymphoma] Lymphoblastoid cells人彌漫大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:1:2-1:3傳代;每周換液2-3次。
MDAMB330細(xì)胞系相關(guān)產(chǎn)品:Sarcoma OSteogenic-2細(xì)胞、OsA-CL細(xì)胞、MADISON LUNG TA-109細(xì)胞
HcerEpic細(xì)胞系相關(guān)產(chǎn)品:H-596細(xì)胞、Hepa1-6細(xì)胞、hUC-MSC細(xì)胞
QGY-7701細(xì)胞系相關(guān)產(chǎn)品:CMT 93細(xì)胞、CAL 12T細(xì)胞、U20S細(xì)胞
CC-LP-1細(xì)胞系相關(guān)產(chǎn)品:HCC-2279細(xì)胞、TE-11細(xì)胞、HEC1B細(xì)胞
58F細(xì)胞系相關(guān)產(chǎn)品:BNL CL2細(xì)胞、H1395細(xì)胞、NCI-H2135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
HBL-1 [Human diffuse large B-cell lymphoma] Lymphoblastoid cells人彌漫大B淋巴瘤細(xì)胞系
細(xì)胞生長(zhǎng)特性:懸浮
細(xì)胞形態(tài)特性:詳見(jiàn)細(xì)胞說(shuō)明書(shū)
為什么培養(yǎng)的細(xì)胞要及時(shí)傳代?體外培養(yǎng)的細(xì)胞大多具有生長(zhǎng)接觸抑制的特性,也就是說(shuō)當(dāng)一個(gè)細(xì)胞被其他細(xì)胞包圍的時(shí)候,它就會(huì)停止生長(zhǎng),及時(shí)傳代后,細(xì)胞的生長(zhǎng)得以繼續(xù)。培養(yǎng)細(xì)胞生長(zhǎng)減慢的原因在哪些?其解決辦法有哪些?可能的原因有:更換了不同的培養(yǎng)液或血清;培養(yǎng)液中一些細(xì)胞生長(zhǎng)必需成分如谷眈肢或生長(zhǎng)因子等耗盡或缺乏或己被破壞;培養(yǎng)物中有少量細(xì)菌或真菌污染;試劑保存不當(dāng);比較新培養(yǎng)液與原培養(yǎng)液成分,比較新血清與舊血清等,找出其它可能的原因。解決辦法:增加起始培養(yǎng)細(xì)胞濃度;讓細(xì)胞逐漸適應(yīng)新培養(yǎng)液;換入新鮮配制培養(yǎng)液;補(bǔ)加谷酷肢或生長(zhǎng)因子等;用無(wú)抗生素培養(yǎng)液培養(yǎng),如發(fā)現(xiàn)污染,去棄培養(yǎng)物,或用抗生素除菌;血清需保存在5℃到20℃; 培養(yǎng)液需在2-8℃避光保存;含血清完全培養(yǎng)液在2-8℃保存,并在2周內(nèi)用完;分離培養(yǎng)物,檢測(cè)支原體。冷凍管應(yīng)如何解凍?取出冷凍管后,須立即放入37℃水槽中快速解凍,輕搖冷凍管使其在1-2分鐘內(nèi)大部分融化,特別注意,需殘留一小塊冰塊,就可拿到超凈工作臺(tái)操作細(xì)胞,用75%酒精消毒凍存管,用新鮮培養(yǎng)基將細(xì)胞轉(zhuǎn)移至培養(yǎng)瓶。另外冷凍管由液氮桶中取出解凍時(shí),必須注意安全,預(yù)防冷凍管爆裂。
BALL1細(xì)胞系相關(guān)產(chǎn)品:Wien 133細(xì)胞、HEH2細(xì)胞、UCLA-SO-M21細(xì)胞
Hs 675.T細(xì)胞系相關(guān)產(chǎn)品:PanC1細(xì)胞、KYSE-270細(xì)胞、NHEK細(xì)胞
OVCAR-4細(xì)胞系相關(guān)產(chǎn)品:WML2細(xì)胞、MBT2細(xì)胞、H-2591細(xì)胞
Medical Research Council cell strain-5細(xì)胞系相關(guān)產(chǎn)品:SupT1細(xì)胞、G 292 Clone A 141B1細(xì)胞、IR983F細(xì)胞
SW1463細(xì)胞系相關(guān)產(chǎn)品:H69細(xì)胞、SF539細(xì)胞、TE1細(xì)胞
SGC996細(xì)胞系相關(guān)產(chǎn)品:HEI-193細(xì)胞、HDF-a細(xì)胞、SNU-407細(xì)胞
NCI-H1373細(xì)胞系相關(guān)產(chǎn)品:HLMVEC細(xì)胞、Colon 38細(xì)胞、RIN Cl-5F細(xì)胞
Centre Antoine Lacassagne-39細(xì)胞系相關(guān)產(chǎn)品:Mv 1 Lu (NBL-7)細(xì)胞、High 5細(xì)胞、CD 18細(xì)胞
TF-1a細(xì)胞系相關(guān)產(chǎn)品:Karpas299細(xì)胞、KYSE-140細(xì)胞、S37細(xì)胞
LC-1/sq細(xì)胞系相關(guān)產(chǎn)品:Panc 10.05細(xì)胞、Scott細(xì)胞、EC-GI細(xì)胞
HBL-1 [Human diffuse large B-cell lymphoma] Lymphoblastoid cells人彌漫大B淋巴瘤細(xì)胞系
OVCAR420細(xì)胞系相關(guān)產(chǎn)品:SiHa細(xì)胞、MDA MB 134VI細(xì)胞、SF767細(xì)胞
Hx147細(xì)胞系相關(guān)產(chǎn)品:343 MG細(xì)胞、BJAB細(xì)胞、COLO 394細(xì)胞
NCIN87細(xì)胞系相關(guān)產(chǎn)品:H1876細(xì)胞、NCIH841細(xì)胞、NCI-H2405細(xì)胞
LLC1細(xì)胞系相關(guān)產(chǎn)品:JHH-7細(xì)胞、MDA-MB435細(xì)胞、JROECL33細(xì)胞
SNU-719細(xì)胞系相關(guān)產(chǎn)品:HcerEpic細(xì)胞、P36細(xì)胞、Sp2/0-Ag14細(xì)胞
YD-15細(xì)胞系相關(guān)產(chǎn)品:Ketr3細(xì)胞、NSC34細(xì)胞、Panc-10.05細(xì)胞
YD-15細(xì)胞系相關(guān)產(chǎn)品:Ketr3細(xì)胞、NSC34細(xì)胞、Panc-10.05細(xì)胞
RCK-8細(xì)胞系相關(guān)產(chǎn)品:University of Arizona Cell Culture-893細(xì)胞、WC00079細(xì)胞、RPMI 7666細(xì)胞
MCA 205細(xì)胞系相關(guān)產(chǎn)品:SW 1417細(xì)胞、Vero-E6細(xì)胞、SCL1細(xì)胞
SCC90細(xì)胞系相關(guān)產(chǎn)品:COLO.205細(xì)胞、U-373MG ATCC細(xì)胞、W256細(xì)胞
SUM-159細(xì)胞系相關(guān)產(chǎn)品:NCI-H1341細(xì)胞、HS5細(xì)胞、JurkatE6-1細(xì)胞
HBL-100細(xì)胞系相關(guān)產(chǎn)品:NS-I/1細(xì)胞、SKMel-28細(xì)胞、Hs27細(xì)胞
H889細(xì)胞系相關(guān)產(chǎn)品:MDAMB415細(xì)胞、MDA.MB.435細(xì)胞、Ramos 2G6 4C10細(xì)胞
Me Wo細(xì)胞系相關(guān)產(chǎn)品:MDAMB453細(xì)胞、SUM 159PT細(xì)胞、SW 1417細(xì)胞
NBL-3細(xì)胞系相關(guān)產(chǎn)品:RL952細(xì)胞、H-2085細(xì)胞、WSUDLCL2細(xì)胞
COLO 201細(xì)胞系相關(guān)產(chǎn)品:Madin Darby Canine Kidney細(xì)胞、HuP-T3細(xì)胞、Bovine Turbinate細(xì)胞
H-441細(xì)胞系相關(guān)產(chǎn)品:769-P細(xì)胞、OVCAR 5細(xì)胞、SUM-190PT細(xì)胞
HSC3細(xì)胞系相關(guān)產(chǎn)品:Panc3_27細(xì)胞、NCI-H498細(xì)胞、HuTu-80細(xì)胞
D341 Med細(xì)胞系相關(guān)產(chǎn)品:A172細(xì)胞、ZR-75-30細(xì)胞、Doubling time: ~50 hours (ATCC).細(xì)胞
H-250細(xì)胞系相關(guān)產(chǎn)品:H520細(xì)胞、Functional Liver Cell-7細(xì)胞、OUMS-27細(xì)胞
RCM1細(xì)胞系相關(guān)產(chǎn)品:J-82細(xì)胞、RMS 1598細(xì)胞、Laboratory of Allergic Diseases 2細(xì)胞
H-378細(xì)胞系相關(guān)產(chǎn)品:SW839細(xì)胞、NCI-H87細(xì)胞、Alpha Mouse Liver 12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
RS(4;11)細(xì)胞系相關(guān)產(chǎn)品:Stanford University Pediatric T-cell line 1細(xì)胞、MUTZ1細(xì)胞、THLE2細(xì)胞
HLFa細(xì)胞系相關(guān)產(chǎn)品:H1563細(xì)胞、SNU-398細(xì)胞、H2196細(xì)胞
HS-766T細(xì)胞系相關(guān)產(chǎn)品:SW-1353細(xì)胞、NCI-H2085細(xì)胞、C32r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
SKUT-1細(xì)胞系相關(guān)產(chǎn)品:H-498細(xì)胞、Kit225細(xì)胞、Ra #1細(xì)胞
SCC-9細(xì)胞系相關(guān)產(chǎn)品:Ls-174-T細(xì)胞、TU 686細(xì)胞、A2780/CP70細(xì)胞
NIE 115細(xì)胞系相關(guān)產(chǎn)品:B16-F10-BL6細(xì)胞、Bac1 2F5細(xì)胞、NCIH1395細(xì)胞
OCILY19細(xì)胞系相關(guān)產(chǎn)品:hFOB 1.19細(xì)胞、NCIH2029細(xì)胞、Walker-256細(xì)胞
OKAC1細(xì)胞系相關(guān)產(chǎn)品:P3JHR1細(xì)胞、Stanford University-Diffuse Histiocytic Lymphoma-10細(xì)胞、MC57細(xì)胞
C-32細(xì)胞系相關(guān)產(chǎn)品:EST81細(xì)胞、Cloudman M3細(xì)胞、UK Pan-1細(xì)胞
SKLU1細(xì)胞系相關(guān)產(chǎn)品:BTI-Tn-5B1-4細(xì)胞、H2085細(xì)胞、CNE2細(xì)胞
NCI-H441細(xì)胞系相關(guān)產(chǎn)品:HCC-366細(xì)胞、MN 60細(xì)胞、DITNC1細(xì)胞
PE/CA-PJ34 (clone C12)細(xì)胞系相關(guān)產(chǎn)品:MOLT 4細(xì)胞、Swiss-3T3細(xì)胞、H-2087細(xì)胞
Daoy細(xì)胞系相關(guān)產(chǎn)品:KMB-17細(xì)胞、UMRC-2細(xì)胞、Rat-2細(xì)胞
CORL23細(xì)胞系相關(guān)產(chǎn)品:DoTc2細(xì)胞、Madin Darby Bovine Kidney細(xì)胞、RF/6A細(xì)胞
CORL23細(xì)胞系相關(guān)產(chǎn)品:DoTc2細(xì)胞、Madin Darby Bovine Kidney細(xì)胞、RF/6A細(xì)胞
NTERA2-D1細(xì)胞系相關(guān)產(chǎn)品:TGBC11TKB細(xì)胞、H378細(xì)胞、BT-549細(xì)胞
FRTL-5細(xì)胞系相關(guān)產(chǎn)品:Hs-675-T細(xì)胞、P3X63Ag8.653細(xì)胞、SO-RB50細(xì)胞
NIH:OVCAR-8細(xì)胞系相關(guān)產(chǎn)品:X63-Ag8.653細(xì)胞、QBI-HEK 293A細(xì)胞、IEC-6細(xì)胞
BCPAP細(xì)胞系相關(guān)產(chǎn)品:NCCIT細(xì)胞、X63-AG 8.653細(xì)胞、COLO-357細(xì)胞
SK-N-DZ細(xì)胞系相關(guān)產(chǎn)品:A-1847細(xì)胞、EC109細(xì)胞、BEL 7405細(xì)胞
HCT-8/V細(xì)胞系相關(guān)產(chǎn)品:DR2R 1610細(xì)胞、B-cell Acute Lymphoblastic Leukemia-1細(xì)胞、TCCSUP細(xì)胞
DSL-6A/C1細(xì)胞系相關(guān)產(chǎn)品:OVCAR-420細(xì)胞、MDA175細(xì)胞、A427細(xì)胞
RBMVEC細(xì)胞系相關(guān)產(chǎn)品:DoTc2 4510細(xì)胞、G401細(xì)胞、IPLB-SF-21-AE細(xì)胞
Intestinal Epithelioid Cell line No. 6細(xì)胞系相關(guān)產(chǎn)品:PG-4細(xì)胞、MESSA-DX5細(xì)胞、RCC4細(xì)胞
NCIH1869細(xì)胞系相關(guān)產(chǎn)品:H2.35細(xì)胞、NCI-H345細(xì)胞、NCI-H-295細(xì)胞
COR-L23細(xì)胞系相關(guān)產(chǎn)品:CF Pac1細(xì)胞、J774A1細(xì)胞、NFS 60細(xì)胞
PZ-HPV-7細(xì)胞系相關(guān)產(chǎn)品:NCI-SNU-216細(xì)胞、174xCEM.T2細(xì)胞、HEK-293FT細(xì)胞
HBL-1 [Human diffuse large B-cell lymphoma] Lymphoblastoid cells人彌漫大B淋巴瘤細(xì)胞系
H378細(xì)胞系相關(guān)產(chǎn)品:McA-RH 8994細(xì)胞、H-929細(xì)胞、293H細(xì)胞
GM15452細(xì)胞系相關(guān)產(chǎn)品:Hs 343.T細(xì)胞、HEK-EBNA細(xì)胞、VP 267細(xì)胞
HIBEpiC細(xì)胞系相關(guān)產(chǎn)品:HEK 293A細(xì)胞、RT 112細(xì)胞、NCI-H676細(xì)胞
OVCAR-4細(xì)胞系相關(guān)產(chǎn)品:WML2細(xì)胞、CT-26細(xì)胞、PLA 802細(xì)胞
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