"NCI-H1770 epithelioid cells人非小細(xì)胞肺癌細(xì)胞系 細(xì)胞背景資料:詳見(jiàn)相關(guān)文獻(xiàn)介紹 細(xì)胞形態(tài):上皮細(xì)胞樣 細(xì)胞復(fù)蘇后貼壁細(xì)胞較少的問(wèn)題分析:總結(jié)1:復(fù)蘇過(guò)程沒(méi)有問(wèn)題,是否是從液氮拿出直接放入溫水,還有培養(yǎng)箱,二氧化碳濃度,培養(yǎng)基、PH值等環(huán)節(jié)。要么加高濃度FBS 15-20%,看看能否幫助貼壁,當(dāng)然也需要考慮血清問(wèn)題,還有確信拿來(lái)的細(xì)胞沒(méi)問(wèn)題??偨Y(jié)2:首先應(yīng)該懷疑凍存,實(shí)際上復(fù)蘇出問(wèn)題的可能非常小,因?yàn)椴僮骱?jiǎn)單,而且死板。1、你凍存的時(shí)候是不是消化的時(shí)間過(guò)長(zhǎng),這是一般人所注意不到的,即使書(shū)上也不講這個(gè)問(wèn)題,太長(zhǎng)的消化時(shí)間會(huì)讓細(xì)胞復(fù)蘇時(shí)失去貼壁能力,表現(xiàn)為先貼后死,原因是在你復(fù)蘇的時(shí)候細(xì)胞已進(jìn)入凋亡程序,不可逆轉(zhuǎn)的死亡。2、你的凍存液好不好,是什么,甘油還是DMSO,質(zhì)量非常重要,否則也會(huì)死亡。3、你的凍存液的量加的是不是太多,ATCC推薦是不超過(guò)7%,大于5%,太多也不好。4、你在凍存的時(shí)候是不是把DMSO混均勻,這個(gè)有一些影響,但不算太大。5、你的凍存是否按部就班,就是所溫度梯度是不是把握嚴(yán)格,很多人容易忘卻這個(gè)事情,因?yàn)檫@個(gè)東西流程長(zhǎng)。6、如果你細(xì)胞污染,你是否能很快看到,我比我的導(dǎo)師能早一天看到污染。從這個(gè)角度講建議去除離心這步。7、你的細(xì)胞在凍存前是否過(guò)密。還有,不贊成孵箱污染這個(gè)概念的,所有在一個(gè)孵箱里的細(xì)胞都污染一個(gè)細(xì)菌的話,這個(gè)細(xì)菌是源于孵箱的,但這不代表孵箱污染,因?yàn)榉跸錈o(wú)論你如何處理都有大量的細(xì)菌,問(wèn)題在操作。每次污染的原因都要盡可能的找,以后就不犯同樣的問(wèn)題,這個(gè)很重要,不能靠猜,否則你就有可能細(xì)胞養(yǎng)絕最后換課題,這個(gè)見(jiàn)得太多了,別不當(dāng)會(huì)事。 JM細(xì)胞系相關(guān)產(chǎn)品:NCI-H929細(xì)胞、Pro-5WgaRI3C細(xì)胞、GM03190A細(xì)胞 X63Ag8.653細(xì)胞系相關(guān)產(chǎn)品:HT144mel細(xì)胞、Leydig細(xì)胞、SW-962細(xì)胞 H-1688細(xì)胞系相關(guān)產(chǎn)品:MDA330細(xì)胞、LLC-PK-1細(xì)胞、Moorfields/Institute of Ophthalmology-Muller 1細(xì)胞 細(xì)胞生長(zhǎng):貼壁 NCI-H1770 epithelioid cells人非小細(xì)胞肺癌細(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) 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:隨細(xì)胞的生長(zhǎng)而換液 UC-3細(xì)胞系相關(guān)產(chǎn)品:HCFB(HCF)細(xì)胞、NCIH650細(xì)胞、DH-82細(xì)胞 HELF細(xì)胞系相關(guān)產(chǎn)品:BAR-T細(xì)胞、MBT-2細(xì)胞、CAL-120細(xì)胞 H6c7細(xì)胞系相關(guān)產(chǎn)品:Y-1細(xì)胞、253JB-V細(xì)胞、LL/2細(xì)胞 HR1K細(xì)胞系相關(guān)產(chǎn)品:JB6 Cl 30-7b細(xì)胞、TM-4細(xì)胞、IB-RS2細(xì)胞 GM03573A細(xì)胞系相關(guān)產(chǎn)品:Panc 5.04細(xì)胞、NOMO1細(xì)胞、LLC-PK1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 NCI-H1770 epithelioid cells人非小細(xì)胞肺癌細(xì)胞系 細(xì)胞生長(zhǎng)特性:懸浮生長(zhǎng) 細(xì)胞形態(tài)特性:詳見(jiàn)細(xì)胞說(shuō)明書(shū) 細(xì)胞培養(yǎng)實(shí)驗(yàn)中常見(jiàn)問(wèn)題總結(jié):1)一般客戶拿到細(xì)胞后,應(yīng)該注意什么?客戶收到細(xì)胞后先不開(kāi)蓋,放在培養(yǎng)箱靜置若干小時(shí)后(看細(xì)胞密度而定)在倒置顯微鏡下觀察細(xì)胞生長(zhǎng)情況,并對(duì)細(xì)胞進(jìn)行不同倍數(shù)拍照(建議受收細(xì)胞后觀察培養(yǎng)基的顏色和是否有漏液情況,顯微鏡下拍細(xì)胞100X,200X各一張),排除細(xì)胞本身污染的情況;收到細(xì)胞未開(kāi)封,出現(xiàn)污染狀況我們負(fù)責(zé)免費(fèi)發(fā)送一株細(xì)胞。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)液吸出,留下10ml培養(yǎng)液繼續(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2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。細(xì)胞消化液建議使用PBS配制,慎用Hanks液配制;2)快遞細(xì)胞多久能到,是寄凍存的細(xì)胞還是復(fù)蘇好的細(xì)胞?我們采用快遞發(fā)貨,一般外地2--3天,寄細(xì)胞前請(qǐng)確認(rèn)當(dāng)?shù)販囟?,如果氣溫低于4度的,則采用郵寄凍存細(xì)胞;3)可否使用與原先培養(yǎng)條件不同之培養(yǎng)基?不能。每一細(xì)胞株均有其特定使用且已適應(yīng)之細(xì)胞培養(yǎng)基,若驟然使用和原先提供之培養(yǎng)條件不同之培養(yǎng)基,細(xì)胞大都無(wú)法立即適應(yīng),造成細(xì)胞無(wú)法存活;4)可否使用與原先培養(yǎng)條件不同之血清種類?不能。血清是細(xì)胞培養(yǎng)上一個(gè)極為重要的營(yíng)養(yǎng)來(lái)源,所以血清的種類和品質(zhì)對(duì)于細(xì)胞的生長(zhǎng)會(huì)產(chǎn)生極大的影響。來(lái)自不同物種的血清,在一些物質(zhì)或分子的量或內(nèi)容物上都有所不同,血清使用錯(cuò)誤常會(huì)造成細(xì)胞無(wú)法存活。 MES-SA/Dx5細(xì)胞系相關(guān)產(chǎn)品:Glioma-261細(xì)胞、B16-F1細(xì)胞、HANK1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 COLO-357細(xì)胞系相關(guān)產(chǎn)品:SNU449細(xì)胞、S91細(xì)胞、MM1.S細(xì)胞 CHOS細(xì)胞系相關(guān)產(chǎn)品:MCF 7細(xì)胞、HBE 135-E6E7細(xì)胞、TYKnu細(xì)胞 Virginia Mason Research Center-Lung Cancer D細(xì)胞系相關(guān)產(chǎn)品:H211細(xì)胞、CLL-CII細(xì)胞、bEnd.3[BEND3]細(xì)胞 CORL26細(xì)胞系相關(guān)產(chǎn)品:WM-239A細(xì)胞、KMS11細(xì)胞、MDAPC2B細(xì)胞 RBE細(xì)胞系相關(guān)產(chǎn)品:IGROV-1細(xì)胞、HEK 293 c18細(xì)胞、NS1-Ag4/1細(xì)胞 UPCISCC154細(xì)胞系相關(guān)產(chǎn)品:FTC133細(xì)胞、SK-N-BE(1)n細(xì)胞、138 MG細(xì)胞 HCCLM3細(xì)胞系相關(guān)產(chǎn)品:HAEC細(xì)胞、Panc_03_27細(xì)胞、EB-3細(xì)胞 OS-732細(xì)胞系相關(guān)產(chǎn)品:6T-CEM細(xì)胞、H-EMC-SS細(xì)胞、M-1細(xì)胞 NCI-H1770 epithelioid cells人非小細(xì)胞肺癌細(xì)胞系 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 BTI-Tn5B14細(xì)胞系相關(guān)產(chǎn)品:Hs 281.T細(xì)胞、CNE2細(xì)胞、LNCaP-ATCC細(xì)胞 J 774A.1細(xì)胞系相關(guān)產(chǎn)品:Panc4.03細(xì)胞、SKCol1細(xì)胞、KBM-5細(xì)胞 NCI-H838細(xì)胞系相關(guān)產(chǎn)品:hTERT-RPE-1細(xì)胞、R1610細(xì)胞、Okayama University Medical School-23細(xì)胞 H2122細(xì)胞系相關(guān)產(chǎn)品:HTR-8/SV-neo細(xì)胞、H2023細(xì)胞、RMa-bm細(xì)胞 COLO-738細(xì)胞系相關(guān)產(chǎn)品:Roswell Park Memorial Institute 1846細(xì)胞、M3 Clone M-3細(xì)胞、GM05862細(xì)胞 COLO-738細(xì)胞系相關(guān)產(chǎn)品:Roswell Park Memorial Institute 1846細(xì)胞、M3 Clone M-3細(xì)胞、GM05862細(xì)胞 CHL-CL-11細(xì)胞系相關(guān)產(chǎn)品:AE1201細(xì)胞、H-2228細(xì)胞、H125細(xì)胞 MOLT-3細(xì)胞系相關(guān)產(chǎn)品:HKBML細(xì)胞、NE1-E6E7細(xì)胞、BEL-7402細(xì)胞 Pt K1細(xì)胞系相關(guān)產(chǎn)品:Ly18細(xì)胞、PZ-HPV-7細(xì)胞、H2452細(xì)胞 Panc-10.05細(xì)胞系相關(guān)產(chǎn)品:OVCA432_Bast細(xì)胞、GP2-293細(xì)胞、Rat1細(xì)胞 HCCLM3細(xì)胞系相關(guān)產(chǎn)品:HAEC細(xì)胞、Panc_03_27細(xì)胞、EB-3細(xì)胞 NCI-H1417細(xì)胞系相關(guān)產(chǎn)品:KTCTL-140細(xì)胞、SNU-620細(xì)胞、HuT78細(xì)胞 OCI AML3細(xì)胞系相關(guān)產(chǎn)品:SW 839細(xì)胞、MOLP-8細(xì)胞、OVISE細(xì)胞 WEHI 231細(xì)胞系相關(guān)產(chǎn)品:Tn 5B1-4細(xì)胞、H1793細(xì)胞、H647細(xì)胞 Balb/c3T3細(xì)胞系相關(guān)產(chǎn)品:IPLB-SF 21AE細(xì)胞、DLM8細(xì)胞、BL6細(xì)胞 GM00215細(xì)胞系相關(guān)產(chǎn)品:OS-RC-2細(xì)胞、NCIH524細(xì)胞、OCI AML2細(xì)胞 DR2R 1610細(xì)胞系相關(guān)產(chǎn)品:GNM細(xì)胞、DLD 1細(xì)胞、Panc2_03細(xì)胞 PANC0504細(xì)胞系相關(guān)產(chǎn)品:ATN1細(xì)胞、HIT細(xì)胞、SK-MEL-24細(xì)胞 BT474細(xì)胞系相關(guān)產(chǎn)品:H-727細(xì)胞、McA-RH8994細(xì)胞、MS-751細(xì)胞 NCIH2009細(xì)胞系相關(guān)產(chǎn)品:FD-LSC-1細(xì)胞、CCD1095Sk細(xì)胞、CT26.WT細(xì)胞 SCCVII細(xì)胞系相關(guān)產(chǎn)品:U-373MG ATCC細(xì)胞、Vx-2細(xì)胞、SiHa細(xì)胞 NPA 87細(xì)胞系相關(guān)產(chǎn)品:L-6TG細(xì)胞、32Dcl3細(xì)胞、Swiss 3T3細(xì)胞 C3H-10T1/2細(xì)胞系相關(guān)產(chǎn)品:MUTZ-3細(xì)胞、ATN1細(xì)胞、Hs 281.T細(xì)胞 U373細(xì)胞系相關(guān)產(chǎn)品:ASPC1細(xì)胞、BIC1細(xì)胞、Y-79細(xì)胞 XuanWei Lung Cancer-05細(xì)胞系相關(guān)產(chǎn)品:GM 2132細(xì)胞、MFE 280細(xì)胞、R D細(xì)胞 H520細(xì)胞系相關(guān)產(chǎn)品:EoL-1 cell細(xì)胞、NCIH660細(xì)胞、MLA-144細(xì)胞 Hs888T細(xì)胞系相關(guān)產(chǎn)品:C4 I細(xì)胞、LPC-H12細(xì)胞、GOS3細(xì)胞 NCIH2172細(xì)胞系相關(guān)產(chǎn)品:MCA-205細(xì)胞、KY-50細(xì)胞、HBMEC細(xì)胞 NCI-H735細(xì)胞系相關(guān)產(chǎn)品:OVCA433細(xì)胞、MES-SA Dx5細(xì)胞、OsACL細(xì)胞 KYSE 270細(xì)胞系相關(guān)產(chǎn)品:HOP92細(xì)胞、Hs739T細(xì)胞、G-361細(xì)胞 Panc-813細(xì)胞系相關(guān)產(chǎn)品:IA-LM細(xì)胞、PCI:SG-231細(xì)胞、OE-19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 Co320細(xì)胞系相關(guān)產(chǎn)品:HOMEC細(xì)胞、H-508細(xì)胞、SU-DHL1細(xì)胞 PVEC細(xì)胞系相關(guān)產(chǎn)品:L-5178-Y細(xì)胞、MDAMB453細(xì)胞、HeLaS3細(xì)胞 CL1.0細(xì)胞系相關(guān)產(chǎn)品:COR-L 105細(xì)胞、M14-MEL細(xì)胞、MMAc-Serum Free細(xì)胞 FHL-124細(xì)胞系相關(guān)產(chǎn)品:SK-MEL3細(xì)胞、SkChA-1細(xì)胞、LUDLU1細(xì)胞 NPC-TW 039細(xì)胞系相關(guān)產(chǎn)品:Leghorn Male Hepatoma cell line細(xì)胞、LLC PK1細(xì)胞、CAL51細(xì)胞 SUPM2細(xì)胞系相關(guān)產(chǎn)品:GM05887A細(xì)胞、JAR細(xì)胞、Tu177細(xì)胞 NTera2/D1細(xì)胞系相關(guān)產(chǎn)品:PK13細(xì)胞、LOUNH91細(xì)胞、HEK293A細(xì)胞 KYSE-50細(xì)胞系相關(guān)產(chǎn)品:Hs604T細(xì)胞、Calu 3細(xì)胞、SW 620細(xì)胞 H1385細(xì)胞系相關(guān)產(chǎn)品:MM.1S細(xì)胞、MD Anderson-Metastatic Breast-175-VIII細(xì)胞、HeLa229細(xì)胞 HSC-3細(xì)胞系相關(guān)產(chǎn)品:NCIH2081細(xì)胞、Nb2細(xì)胞、HUCEC細(xì)胞 MV411細(xì)胞系相關(guān)產(chǎn)品:SCC7細(xì)胞、Transformed C3H Mouse Kidney-1細(xì)胞、PFSK細(xì)胞 Michigan Cancer Foundation-12A細(xì)胞系相關(guān)產(chǎn)品:NBL-12細(xì)胞、HSC/mHSC細(xì)胞、CAL27細(xì)胞 HL-7702細(xì)胞系相關(guān)產(chǎn)品:Ly3細(xì)胞、H508細(xì)胞、BPH1細(xì)胞 K7M2細(xì)胞系相關(guān)產(chǎn)品:GM03569D細(xì)胞、Pro-Lec1.3C細(xì)胞、NCI-H1092細(xì)胞 IOSE-80細(xì)胞系相關(guān)產(chǎn)品:mouse Inner Medullary Collecting Duct-3細(xì)胞、LO2細(xì)胞、H1781細(xì)胞 HCT8細(xì)胞系相關(guān)產(chǎn)品:P3 NS1 Ag4細(xì)胞、G361mel細(xì)胞、MDA MB231細(xì)胞 PLC PRF 5細(xì)胞系相關(guān)產(chǎn)品:RLE-6TN細(xì)胞、U-2932細(xì)胞、DMS-53細(xì)胞 SPDC-CCL141細(xì)胞系相關(guān)產(chǎn)品:CCC-HEH-2細(xì)胞、CHP-126細(xì)胞、LTEP a2細(xì)胞 NCI-H1770 epithelioid cells人非小細(xì)胞肺癌細(xì)胞系 PC-12細(xì)胞系相關(guān)產(chǎn)品:SW-13細(xì)胞、P3X63NS1細(xì)胞、KM933細(xì)胞 NU-GC-4細(xì)胞系相關(guān)產(chǎn)品:TK-1細(xì)胞、KYSE0030細(xì)胞、NBL-1細(xì)胞 DHBE細(xì)胞系相關(guān)產(chǎn)品:SW900細(xì)胞、NCI-H165細(xì)胞、108CC15細(xì)胞 COLO-357細(xì)胞系相關(guān)產(chǎn)品:SNU449細(xì)胞、BS-C-1細(xì)胞、H2291細(xì)胞 "
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