"MFC epithelioid cells小鼠胃癌細(xì)胞系 細(xì)胞背景資料:胃癌 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。 NS-20Y細(xì)胞系相關(guān)產(chǎn)品:CCRF CEM細(xì)胞、VK2細(xì)胞、U2932細(xì)胞 HCC-2218細(xì)胞系相關(guān)產(chǎn)品:WRL 68細(xì)胞、SCC4細(xì)胞、RT-BM 1細(xì)胞 Caki-1細(xì)胞系相關(guān)產(chǎn)品:NPA-87細(xì)胞、NCIH929細(xì)胞、GM-3573細(xì)胞 細(xì)胞生長(zhǎng):貼壁 MFC epithelioid cells小鼠胃癌細(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:1:2-1:3傳代;每周換液2-3次。 WIL2 Secreting細(xì)胞系相關(guān)產(chǎn)品:H-441細(xì)胞、SNU-1細(xì)胞、Human fetal lung fibroblast 1細(xì)胞 MOLP-2細(xì)胞系相關(guān)產(chǎn)品:NCI-H157細(xì)胞、H-510A細(xì)胞、HCC-2935細(xì)胞 HCC44細(xì)胞系相關(guān)產(chǎn)品:NCIH358細(xì)胞、SF295細(xì)胞、ME1細(xì)胞 SW-982細(xì)胞系相關(guān)產(chǎn)品:LADMAC細(xì)胞、C4I細(xì)胞、YD-38細(xì)胞 HCa/16A3-F細(xì)胞系相關(guān)產(chǎn)品:SHEP1細(xì)胞、CCK-81細(xì)胞、RA.1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 MFC epithelioid cells小鼠胃癌細(xì)胞系 細(xì)胞生長(zhǎng)特性:貼壁 細(xì)胞形態(tài)特性:詳見(jiàn)細(xì)胞說(shuō)明書(shū) 購(gòu)買(mǎi)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。 MN-60細(xì)胞系相關(guān)產(chǎn)品:Wien133細(xì)胞、H-838細(xì)胞、HCC1599細(xì)胞 HCA 7細(xì)胞系相關(guān)產(chǎn)品:H-292細(xì)胞、GC1-SPG細(xì)胞、AR4-IP細(xì)胞 SUDHL-5細(xì)胞系相關(guān)產(chǎn)品:MX-1細(xì)胞、NCI-H929細(xì)胞、SKml2細(xì)胞 Shadyside Hospital Pittsburgh-77細(xì)胞系相關(guān)產(chǎn)品:H9c2 (2-1)細(xì)胞、R1800[RA]細(xì)胞、M059K細(xì)胞 526mel細(xì)胞系相關(guān)產(chǎn)品:MC3T3-E1(C4)細(xì)胞、RPMI7951細(xì)胞、HD11細(xì)胞 H209細(xì)胞系相關(guān)產(chǎn)品:HN6細(xì)胞、SKHEP1細(xì)胞、SPCA1細(xì)胞 L 428細(xì)胞系相關(guān)產(chǎn)品:SNU475細(xì)胞、HCC-1833細(xì)胞、QGP1細(xì)胞 SW626細(xì)胞系相關(guān)產(chǎn)品:XWLC-05細(xì)胞、VMRC-LCD細(xì)胞、HCV29細(xì)胞 H-727細(xì)胞系相關(guān)產(chǎn)品:293A細(xì)胞、U251細(xì)胞、PK-13細(xì)胞 LA-N-5細(xì)胞系相關(guān)產(chǎn)品:Merwin Plasma Cell tumor-11細(xì)胞、NIH:OVCAR-8細(xì)胞、LAD-2細(xì)胞 MFC epithelioid cells小鼠胃癌細(xì)胞系 Hs 274.T細(xì)胞系相關(guān)產(chǎn)品:KU.812細(xì)胞、HECV細(xì)胞、NCIH23細(xì)胞 HEK 293A細(xì)胞系相關(guān)產(chǎn)品:ESC-410細(xì)胞、LC-1/sq細(xì)胞、RGC5細(xì)胞 Mahlavu細(xì)胞系相關(guān)產(chǎn)品:Sp2/0-Ag14細(xì)胞、MB468細(xì)胞、HO-1-N-1細(xì)胞 L78細(xì)胞系相關(guān)產(chǎn)品:L929(NCTC)細(xì)胞、HEC-151細(xì)胞、MCF-12F細(xì)胞 T/G HA-VSMC細(xì)胞系相關(guān)產(chǎn)品:OVCAR-433細(xì)胞、PLC/PRF5細(xì)胞、NCI-H1563細(xì)胞 ECV細(xì)胞系相關(guān)產(chǎn)品:S91細(xì)胞、NCIH522細(xì)胞、H1954細(xì)胞 ECV細(xì)胞系相關(guān)產(chǎn)品:S91細(xì)胞、NCIH522細(xì)胞、H1954細(xì)胞 JKT-1細(xì)胞系相關(guān)產(chǎn)品:HEYA8細(xì)胞、NCI-SNU-449細(xì)胞、HCC0366細(xì)胞 HEK293-EBNA1細(xì)胞系相關(guān)產(chǎn)品:16HBE細(xì)胞、M21細(xì)胞、IPECJ2細(xì)胞 H128細(xì)胞系相關(guān)產(chǎn)品:Caco2BBe細(xì)胞、MBVP細(xì)胞、MGH-U3 (RN)細(xì)胞 SK-MEL2細(xì)胞系相關(guān)產(chǎn)品:COLO824細(xì)胞、NCIH2286細(xì)胞、NBL-2細(xì)胞 PAEC細(xì)胞系相關(guān)產(chǎn)品:HCT-8/V細(xì)胞、293/EBNA-1細(xì)胞、2B4-L細(xì)胞 SUM102細(xì)胞系相關(guān)產(chǎn)品:CPAE細(xì)胞、GS9L細(xì)胞、H2.35細(xì)胞 Hela-mock細(xì)胞系相關(guān)產(chǎn)品:LS-123細(xì)胞、HCV 29細(xì)胞、Hs 578.Bst細(xì)胞 PL-45細(xì)胞系相關(guān)產(chǎn)品:KPNRTBM1細(xì)胞、SAOS 2細(xì)胞、DHL-4細(xì)胞 EFO-27細(xì)胞系相關(guān)產(chǎn)品:PLA-802細(xì)胞、NCIH1092細(xì)胞、PNT1A細(xì)胞 MMAc.SF細(xì)胞系相關(guān)產(chǎn)品:PIEC細(xì)胞、NCIH2452細(xì)胞、CDC/EU.HMEC-1細(xì)胞 KM932細(xì)胞系相關(guān)產(chǎn)品:T-24細(xì)胞、Caco-2細(xì)胞、HRC-99細(xì)胞 BLO-11細(xì)胞系相關(guān)產(chǎn)品:3T3-Swiss細(xì)胞、MCF 7B細(xì)胞、16HBE細(xì)胞 U-hth74細(xì)胞系相關(guān)產(chǎn)品:MEL-526細(xì)胞、HuT-78細(xì)胞、P3HR1細(xì)胞 Rat Glomerular Endothelial細(xì)胞系相關(guān)產(chǎn)品:NCIH747細(xì)胞、JeKo-1細(xì)胞、NCI747細(xì)胞 TMK1細(xì)胞系相關(guān)產(chǎn)品:HMEC細(xì)胞、NCIH1819細(xì)胞、RSC-96細(xì)胞 KAT5細(xì)胞系相關(guān)產(chǎn)品:HTori-3.1細(xì)胞、MHH-CALL2細(xì)胞、H-1404細(xì)胞 Tb 1 Lu (NBL-12)細(xì)胞系相關(guān)產(chǎn)品:VMM-5A細(xì)胞、SU-DHL-2細(xì)胞、RASMCs細(xì)胞 Acanthosis Nigricans 3rd attempt-CArcinoma細(xì)胞系相關(guān)產(chǎn)品:U251細(xì)胞、SNU1040細(xì)胞、MLFC細(xì)胞 L23/P細(xì)胞系相關(guān)產(chǎn)品:ETCC007細(xì)胞、Mink細(xì)胞、Primary Liver Carcinoma/Poliomyelitis Research Foundation/5細(xì)胞 Madin-Darby Canine Kidney細(xì)胞系相關(guān)產(chǎn)品:U-251細(xì)胞、CCC-HEL-1細(xì)胞、RERF-LC-MS細(xì)胞 Hs606T細(xì)胞系相關(guān)產(chǎn)品:FL-83B細(xì)胞、H322細(xì)胞、HCC2935細(xì)胞 WERI-Rb 1細(xì)胞系相關(guān)產(chǎn)品:C4-2細(xì)胞、RK13細(xì)胞、Y3-Ag1,2,3細(xì)胞 MGc80-3細(xì)胞系相關(guān)產(chǎn)品:Hep 3B細(xì)胞、T98-G細(xì)胞、OVCAR432細(xì)胞 LN382細(xì)胞系相關(guān)產(chǎn)品:526mel細(xì)胞、PL5細(xì)胞、GC1-SPG細(xì)胞 MM.1S細(xì)胞系相關(guān)產(chǎn)品:HCC-1419細(xì)胞、C-4I細(xì)胞、F36P細(xì)胞 293GP細(xì)胞系相關(guān)產(chǎn)品:BT.549細(xì)胞、H-23細(xì)胞、R3/1細(xì)胞 LIM-1215細(xì)胞系相關(guān)產(chǎn)品:PANC0327細(xì)胞、CEMx721.174.T2細(xì)胞、GM00637F細(xì)胞 H322T細(xì)胞系相關(guān)產(chǎn)品:H676B細(xì)胞、MV-522細(xì)胞、U138MG細(xì)胞 MDA-MB-468-RED細(xì)胞系相關(guān)產(chǎn)品:GM346細(xì)胞、Medical Research Council cell strain-5細(xì)胞、Me Wo細(xì)胞 TE4細(xì)胞系相關(guān)產(chǎn)品:RKO_AS45細(xì)胞、SNB-19細(xì)胞、BrCL18細(xì)胞 MV4II細(xì)胞系相關(guān)產(chǎn)品:Asian Medical Center-Head and Neck cancer-8細(xì)胞、KYSE520細(xì)胞、HFLS-RA細(xì)胞 HOPC細(xì)胞系相關(guān)產(chǎn)品:UACC-893細(xì)胞、Normal Rat Kidney細(xì)胞、H209細(xì)胞 Tca-83細(xì)胞系相關(guān)產(chǎn)品:Centre Antoine Lacassagne-33細(xì)胞、NCI-H1648細(xì)胞、NCI-H1355細(xì)胞 OVCA-433細(xì)胞系相關(guān)產(chǎn)品:HOS細(xì)胞、NCC-IT細(xì)胞、Bowes melanoma cells細(xì)胞 EC-109細(xì)胞系相關(guān)產(chǎn)品:HRIF細(xì)胞、P3-X63Ag8.653細(xì)胞、SUDHL-1細(xì)胞 GM03320D細(xì)胞系相關(guān)產(chǎn)品:TK.10細(xì)胞、HLEB3細(xì)胞、RH35細(xì)胞 MLE-12細(xì)胞系相關(guān)產(chǎn)品:MDA361細(xì)胞、Sp 2817細(xì)胞、TMK1細(xì)胞 MLMA細(xì)胞系相關(guān)產(chǎn)品:NCI-H727細(xì)胞、COLO-16細(xì)胞、SW1417細(xì)胞 Bx-PC3細(xì)胞系相關(guān)產(chǎn)品:Hs832T細(xì)胞、HOMEC細(xì)胞、L78細(xì)胞 P-388細(xì)胞系相關(guān)產(chǎn)品:ACC-3細(xì)胞、SKNBE2細(xì)胞、HLF細(xì)胞 U251細(xì)胞系相關(guān)產(chǎn)品:Human Lung Microvascular Endothelial Cell line-5a細(xì)胞、H-2444細(xì)胞、B16-F10-BL6細(xì)胞 HEI193細(xì)胞系相關(guān)產(chǎn)品:DLD1細(xì)胞、138MG細(xì)胞、UMNSAH/DF-1細(xì)胞 HEK293-A細(xì)胞系相關(guān)產(chǎn)品:UPCI:SCC154細(xì)胞、H-2170細(xì)胞、C-4-I細(xì)胞 NCIH2228細(xì)胞系相關(guān)產(chǎn)品:KCL-22細(xì)胞、FHL-124細(xì)胞、ZYM-SVEC01細(xì)胞 MFC epithelioid cells小鼠胃癌細(xì)胞系 FCCH1018細(xì)胞系相關(guān)產(chǎn)品:VP 229細(xì)胞、210RCY3-Ag1.2.3細(xì)胞、EFM-192C細(xì)胞 Okayama University Medical School-27細(xì)胞系相關(guān)產(chǎn)品:EHEB細(xì)胞、JTC-28細(xì)胞、4T1-A細(xì)胞 WEHI164細(xì)胞系相關(guān)產(chǎn)品:RMS 13細(xì)胞、MDA-MB436細(xì)胞、BC-3-H-I細(xì)胞 SUDHL-5細(xì)胞系相關(guān)產(chǎn)品:MX-1細(xì)胞、MC3T3E1細(xì)胞、SNU-407細(xì)胞 "
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