"4T1 epithelioid cells小鼠乳腺癌細(xì)胞系 細(xì)胞背景資料:4T1是從410.4瘤株中未經(jīng)誘變篩得的6-鳥(niǎo)嘌噙抗性細(xì)胞株。當(dāng)注射到BALB/c小鼠中時(shí),4T1自發(fā)產(chǎn)生高轉(zhuǎn)移腫瘤,可轉(zhuǎn)移到肺,肝,淋巴結(jié)和大腦,同時(shí)在注射部位形成始發(fā)灶。誘導(dǎo)轉(zhuǎn)移時(shí)不需要摘除始發(fā)灶。4T1細(xì)胞在BALB/c小鼠中的生長(zhǎng)與轉(zhuǎn)移特性與人體中的乳腺癌十分相近。這種腫瘤是人VI期乳腺癌的動(dòng)物模型。4T1-誘導(dǎo)的腫瘤在手術(shù)后及未手術(shù)情況下轉(zhuǎn)移的動(dòng)力學(xué)相近,可以用作手術(shù)后及未手術(shù)模型。跟其他腫瘤模型相比,由于4T1的抗6-鳥(niǎo)嘌噙特性,微小的轉(zhuǎn)移細(xì)胞團(tuán)(少到僅僅1個(gè))也可以在許多遠(yuǎn)端器官中檢測(cè)到。沒(méi)必要數(shù)淋巴結(jié)或稱重器官。 細(xì)胞形態(tài):上皮細(xì)胞樣 細(xì)胞株(系)的使用,為醫(yī)學(xué)研究和測(cè)試工作帶來(lái)了極大的方便。但細(xì)胞的傳代是有限制的,長(zhǎng)期連續(xù)傳代的細(xì)胞,不僅消耗大量的人力和物力,而且細(xì)胞的生長(zhǎng)與形態(tài)等會(huì)有一定退變或轉(zhuǎn)化,因而細(xì)胞失去原有的遺傳特性,有時(shí)還會(huì)由于細(xì)胞污染而造成傳代中斷,種子丟失。因此,在實(shí)際工作中常需凍存一定數(shù)量的細(xì)胞,以備替換使用。細(xì)胞冷凍與復(fù)蘇是細(xì)胞培養(yǎng) 室的常規(guī)工作和通用技術(shù)。目前,細(xì)胞凍存最常用的技術(shù)是液氮冷凍保存法,主要采用加適量保護(hù)劑的緩慢冷凍法凍存細(xì)胞。細(xì)胞在不加任何保護(hù)劑的情況下直接冷凍,細(xì)胞內(nèi)外的水分會(huì)很快形成冰晶,從而引起一系列不良反應(yīng)。如細(xì)胞脫水使局部電解質(zhì)濃度增高,pH值改變,部分蛋白質(zhì)由于上述原因而變性,引起細(xì)胞內(nèi)部空間結(jié)構(gòu)紊亂,溶酶體膜由此遭到損傷而釋放出溶酶體酶,使細(xì)胞內(nèi)結(jié)構(gòu)成分造成破壞,線粒體腫脹,功能丟失,并造成能量代謝障礙。胞膜上的類脂蛋白復(fù)合體也易破壞引起細(xì)胞膜通透性的改變,使細(xì)胞內(nèi)容物丟失。如果細(xì)胞內(nèi)冰晶形成較多,隨冷凍溫度的降低,冰晶體積膨脹造成細(xì)胞核DNA空間構(gòu)型發(fā)生不可逆的損傷,而致細(xì)胞死亡。因此,細(xì)胞冷凍技術(shù)的關(guān)鍵是盡可能地減少細(xì)胞內(nèi)水分,減少細(xì)胞內(nèi)冰晶的形成。采用甘油或二甲基亞砜作保護(hù)劑,這兩種物質(zhì)分子量小,溶解度大,易穿透細(xì)胞,可以使冰點(diǎn)下降,提高細(xì)胞膜對(duì)水的通透性,且對(duì)細(xì)胞無(wú)明顯毒性。慢速冷凍方法又可使細(xì)胞內(nèi)的水分滲出細(xì)胞外,減少胞內(nèi)形成冰結(jié)晶的機(jī)會(huì),從而減少冰晶對(duì)細(xì)胞的損傷。 MEL細(xì)胞系相關(guān)產(chǎn)品:RS411細(xì)胞、QGP1細(xì)胞、TE 85 ClF-5細(xì)胞 EC109細(xì)胞系相關(guān)產(chǎn)品:Ra No. 1細(xì)胞、Institute for Medical Research-90細(xì)胞、COLO678細(xì)胞 PL-5細(xì)胞系相關(guān)產(chǎn)品:CCRF-CEM細(xì)胞、UACC-893細(xì)胞、NTera2 cl.D1細(xì)胞 細(xì)胞生長(zhǎng):貼壁 4T1 epithelioid cells小鼠乳腺癌細(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) 細(xì)胞傳代的一般方法:開(kāi)始細(xì)胞傳代前,仔細(xì)檢查細(xì)胞傳代所需的儀器和試劑是否齊全:一般主要有:細(xì)胞(單層細(xì)胞,鏡檢適合)、培養(yǎng)液(MEM-0.2%LH培養(yǎng)液或MEM培養(yǎng)液或199等適合細(xì)胞有要求的培養(yǎng)液)、滅活小牛血清、慶大霉素溶液(1萬(wàn)單位)、7.5%NaHC03溶液、0.25%胰蛋白酶、PBS洗液。用具:小方瓶(100m1)、克氏瓶、膠塞、吸管(10ml、1m1)、細(xì)胞吹打管(20ml)(以上用具需經(jīng)121℃至少15分鐘高壓滅菌)、C02 孵箱、超凈臺(tái)、倒置顯微鏡、4℃、-20℃冰箱;操作步驟:鏡檢細(xì)胞,挑選生長(zhǎng)狀態(tài)良好,細(xì)胞界限清晰,具有立體感,形態(tài)好無(wú)污染、無(wú)異常及可疑病變細(xì)胞。配制細(xì)胞培養(yǎng)液:按以下配比配制MEM-0.2%LH培養(yǎng)液100ml內(nèi),加入滅活小牛血清10m1,慶大霉素溶液0.3m1,7.5%碳酸鈉溶液3ml。(僅供一般參考)。消化細(xì)胞;先用PBS洗液沖洗細(xì)胞表面1-2次,每次10m1,棄去洗液,向細(xì)胞瓶?jī)?nèi)加入0.25%胰蛋白酶溶液,每瓶1m1,細(xì)胞瓶平放,使消化液完全覆蓋細(xì)胞表面。至細(xì)胞完全脫落到胰酶中。每瓶加入10m1細(xì)胞培養(yǎng)液吹打分散細(xì)胞,按所需的傳代比例分裝于小方瓶(100m1)中,再加入10m1細(xì)胞培養(yǎng)液重復(fù)吹打一次后分裝,然后補(bǔ)加至所需培養(yǎng)量。加塞,置于37±1℃孵箱培養(yǎng)。約2~4天成片。(由細(xì)胞接種濃度決定);填寫(xiě)傳代記錄。 細(xì)胞傳代方法:消化3-5分鐘,1:2,3天內(nèi)可長(zhǎng)滿 OV1-P細(xì)胞系相關(guān)產(chǎn)品:H740細(xì)胞、SK-ChA-1細(xì)胞、UPCI:SCC154細(xì)胞 Mv 1 Lu細(xì)胞系相關(guān)產(chǎn)品:Sf-21細(xì)胞、C4-1細(xì)胞、RL-65細(xì)胞 NCIH1373細(xì)胞系相關(guān)產(chǎn)品:H774細(xì)胞、BAR-T細(xì)胞、OVTOKO細(xì)胞 MSTO-211H細(xì)胞系相關(guān)產(chǎn)品:H-748細(xì)胞、Bx-PC3細(xì)胞、ECGI10細(xì)胞 HLF-a細(xì)胞系相關(guān)產(chǎn)品:TW01細(xì)胞、MDAMB330細(xì)胞、COLO.205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 4T1 epithelioid cells小鼠乳腺癌細(xì)胞系 細(xì)胞生長(zhǎng)特性:貼壁生長(zhǎng) 細(xì)胞形態(tài)特性:上皮樣 購(gòu)買的細(xì)胞死亡或細(xì)胞存活率不佳可能原因?研究人員在細(xì)胞培養(yǎng)時(shí)出現(xiàn)存活率不佳,原因比較復(fù)雜,常見(jiàn)原因可歸納為:培養(yǎng)基使用錯(cuò)誤或培養(yǎng)基品質(zhì)不佳;血清使用錯(cuò)誤或血清的品質(zhì)不佳;解凍過(guò)程錯(cuò)誤;冷凍細(xì)胞解凍后,加以洗滌細(xì)胞和離心;懸渾細(xì)胞誤認(rèn)為死細(xì)胞;培養(yǎng)溫度使用錯(cuò)誤;細(xì)胞置于-80℃太久等。建議嚴(yán)格參照ATCC的標(biāo)準(zhǔn)操作規(guī)程進(jìn)行細(xì)胞復(fù)蘇、凍存等工作。欲將一般動(dòng)物細(xì)胞離心下來(lái),其離心速率應(yīng)為多少轉(zhuǎn)速?欲回收動(dòng)物細(xì)胞,其離心速率一般為300×g(約1O(jiān)OOrpm),5-10分鐘,轉(zhuǎn)速過(guò)高或離心時(shí)間過(guò)長(zhǎng)都將造成細(xì)胞死亡。合適的離心轉(zhuǎn)速是根據(jù)相對(duì)離心力決定。RCF=1.119×105×r×(rpm)2,其中r為離心機(jī)轉(zhuǎn)軸中心與離心套管底部?jī)?nèi)壁的距離;rpm為離心機(jī)每分鐘的轉(zhuǎn)數(shù);RCF(relative centrifugal force)為相對(duì)離心力,以重力加速度g(980.66cm/s2)的倍數(shù)來(lái)表示單位。 LS174細(xì)胞系相關(guān)產(chǎn)品:HIBEC細(xì)胞、33604細(xì)胞、H1838細(xì)胞 SKOV-3細(xì)胞系相關(guān)產(chǎn)品:MOLT 3細(xì)胞、MADB-106細(xì)胞、G-292細(xì)胞 Hep3B細(xì)胞系相關(guān)產(chǎn)品:P3J-HR-1細(xì)胞、OCI-AML3細(xì)胞、IPI-2I細(xì)胞 HS-766-T細(xì)胞系相關(guān)產(chǎn)品:HuCCT-1細(xì)胞、COLO 394細(xì)胞、ONS-76細(xì)胞 Adult Retinal Pigment Epithelial cell line-19細(xì)胞系相關(guān)產(chǎn)品:Mono Mac 6細(xì)胞、HOS-MNNG細(xì)胞、LN-229細(xì)胞 COLO-679細(xì)胞系相關(guān)產(chǎn)品:TE-6細(xì)胞、H283細(xì)胞、3T3-442A細(xì)胞 HCC-1954BL細(xì)胞系相關(guān)產(chǎn)品:Walker256-TC細(xì)胞、WSUDLCL2細(xì)胞、MEF細(xì)胞 MKN 7細(xì)胞系相關(guān)產(chǎn)品:MCAEC細(xì)胞、COR-L279細(xì)胞、JEG-3細(xì)胞 H-740細(xì)胞系相關(guān)產(chǎn)品:UACC 812細(xì)胞、H-1436細(xì)胞、HNE-2細(xì)胞 UMUC14細(xì)胞系相關(guān)產(chǎn)品:Mono Mac 1細(xì)胞、CT26-clone 25細(xì)胞、MNNG-HOS細(xì)胞 4T1 epithelioid cells小鼠乳腺癌細(xì)胞系 U-226AR1細(xì)胞系相關(guān)產(chǎn)品:H-1417細(xì)胞、U-937細(xì)胞、WML2細(xì)胞 NCI-H3255細(xì)胞系相關(guān)產(chǎn)品:CATH-a細(xì)胞、C8166-CD4細(xì)胞、H-196細(xì)胞 786.O細(xì)胞系相關(guān)產(chǎn)品:NCIH2347細(xì)胞、Caov-3細(xì)胞、NCIH1838細(xì)胞 alpha TC1.6細(xì)胞系相關(guān)產(chǎn)品:HO1N1細(xì)胞、MIN-6細(xì)胞、BC-024細(xì)胞 H-87細(xì)胞系相關(guān)產(chǎn)品:Panc813細(xì)胞、Karpas299細(xì)胞、ZR75-30細(xì)胞 NE-4C細(xì)胞系相關(guān)產(chǎn)品:PK-136細(xì)胞、rRMECs細(xì)胞、NuTu-19細(xì)胞 NE-4C細(xì)胞系相關(guān)產(chǎn)品:PK-136細(xì)胞、rRMECs細(xì)胞、NuTu-19細(xì)胞 MSC細(xì)胞系相關(guān)產(chǎn)品:CALU1細(xì)胞、GT38細(xì)胞、ARPE-19細(xì)胞 H-2228細(xì)胞系相關(guān)產(chǎn)品:VB細(xì)胞、KE-39細(xì)胞、KP-N-RT-BM細(xì)胞 SNU-484細(xì)胞系相關(guān)產(chǎn)品:T.T細(xì)胞、KTC-1細(xì)胞、EBNA293細(xì)胞 Cates-1B細(xì)胞系相關(guān)產(chǎn)品:BHK 21細(xì)胞、AAV-293細(xì)胞、DC 2.4細(xì)胞 NCIH520細(xì)胞系相關(guān)產(chǎn)品:SUDHL-16細(xì)胞、A 172細(xì)胞、DC2.4細(xì)胞 Neukoplast細(xì)胞系相關(guān)產(chǎn)品:HKF細(xì)胞、Laboratory of Allergic Diseases 2細(xì)胞、TOV21G細(xì)胞 MC-116細(xì)胞系相關(guān)產(chǎn)品:SBC3細(xì)胞、NPA87-1細(xì)胞、H-510細(xì)胞 ZR-75-1細(xì)胞系相關(guān)產(chǎn)品:H-522細(xì)胞、G-Olig2細(xì)胞、DMS-114細(xì)胞 NU-GC-4細(xì)胞系相關(guān)產(chǎn)品:TK-1細(xì)胞、Ramos G6.C10細(xì)胞、BERH-2細(xì)胞 NCI-H2087細(xì)胞系相關(guān)產(chǎn)品:SW962細(xì)胞、2B4細(xì)胞、KYSE30細(xì)胞 HS-294細(xì)胞系相關(guān)產(chǎn)品:KPL-4細(xì)胞、RIN-m5F細(xì)胞、Huh 7.5細(xì)胞 CEM T4細(xì)胞系相關(guān)產(chǎn)品:DHL5細(xì)胞、Douglas Foster-1細(xì)胞、NPA 87細(xì)胞 BE2-M17細(xì)胞系相關(guān)產(chǎn)品:COLO680N細(xì)胞、CDC/EU.HMEC-1細(xì)胞、SW1222細(xì)胞 BCaP 37細(xì)胞系相關(guān)產(chǎn)品:LA-N-1細(xì)胞、PC615.3細(xì)胞、AG06814-N細(xì)胞 RPE-1細(xì)胞系相關(guān)產(chǎn)品:Human Hepatocyte Line 5細(xì)胞、H-1793細(xì)胞、HFLS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 SW 480細(xì)胞系相關(guān)產(chǎn)品:Balb/c 3T3細(xì)胞、CAL120細(xì)胞、NCI/ADR-RES細(xì)胞 PLC-PRF-5細(xì)胞系相關(guān)產(chǎn)品:NCIH345細(xì)胞、DU 4475細(xì)胞、207細(xì)胞 SUDHL-4細(xì)胞系相關(guān)產(chǎn)品:NCI H345細(xì)胞、NW-MEL-38細(xì)胞、H-676B細(xì)胞 Leukemic 1210細(xì)胞系相關(guān)產(chǎn)品:NCI-H2108細(xì)胞、H-1693細(xì)胞、Co320細(xì)胞 NCTC 3960細(xì)胞系相關(guān)產(chǎn)品:MD Anderson-Metastatic Breast-157細(xì)胞、HOS/MNNG細(xì)胞、JAR細(xì)胞 Human Embryonic Skin細(xì)胞系相關(guān)產(chǎn)品:BeWo細(xì)胞、UMUC3細(xì)胞、SUDHL1細(xì)胞 JVM-2細(xì)胞系相關(guān)產(chǎn)品:3T3-L1 ad細(xì)胞、Caco-2 BBe細(xì)胞、SaOS-2細(xì)胞 HMEL細(xì)胞系相關(guān)產(chǎn)品:Strain V細(xì)胞、HEK 293A細(xì)胞、H1184細(xì)胞 149PT細(xì)胞系相關(guān)產(chǎn)品:MOLT-4細(xì)胞、NCIH2405細(xì)胞、MD Anderson-Metastatic Breast-435細(xì)胞 Dysplastic Oral Keratinocyte細(xì)胞系相關(guān)產(chǎn)品:ACC2細(xì)胞、SK-RC-42細(xì)胞、SKMEL1細(xì)胞 NCI-H2106細(xì)胞系相關(guān)產(chǎn)品:Institute for Medical Research-32細(xì)胞、SK-OV-433細(xì)胞、VMM-5A細(xì)胞 YD15細(xì)胞系相關(guān)產(chǎn)品:Hs940T細(xì)胞、Emory University-2細(xì)胞、SKNO-1細(xì)胞 TOV-112D細(xì)胞系相關(guān)產(chǎn)品:ECGI10細(xì)胞、HEK293-H細(xì)胞、Hs68細(xì)胞 BALB/3T3 cl. 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