"Sol8 epithelioid cells小鼠骨骼肌肌肉母細(xì)胞系 細(xì)胞背景資料:骨骼肌;C3H 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。 HCC-1419細(xì)胞系相關(guān)產(chǎn)品:Adeno 293細(xì)胞、MCFs細(xì)胞、INS-1E細(xì)胞 NCI H226細(xì)胞系相關(guān)產(chǎn)品:Human Microvascular Endothelial Cell line-1細(xì)胞、HCC78細(xì)胞、BAR-T細(xì)胞 HS-294-T細(xì)胞系相關(guān)產(chǎn)品:HSAEC1-KT細(xì)胞、TC-1[JHU-1]細(xì)胞、TALL1細(xì)胞 細(xì)胞生長(zhǎng):貼壁 Sol8 epithelioid cells小鼠骨骼肌肌肉母細(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);填寫傳代記錄。 細(xì)胞傳代方法:1:2-1:3傳代;每周換液2-3次。 COR-L23/P細(xì)胞系相關(guān)產(chǎn)品:RKO-E6細(xì)胞、AtT 20細(xì)胞、NALM-6細(xì)胞 LS 174T細(xì)胞系相關(guān)產(chǎn)品:HS 445T細(xì)胞、P3-NS1/1-Ag4-1細(xì)胞、L 540細(xì)胞 C-8161細(xì)胞系相關(guān)產(chǎn)品:PLB-985細(xì)胞、BS-C-1細(xì)胞、SNU-423細(xì)胞 IM95細(xì)胞系相關(guān)產(chǎn)品:CORL279細(xì)胞、Nakata-1細(xì)胞、Panc_04_03細(xì)胞 MKN7細(xì)胞系相關(guān)產(chǎn)品:Panc 1細(xì)胞、HS-445細(xì)胞、WM266mel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 Sol8 epithelioid cells小鼠骨骼肌肌肉母細(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。 Vero-76細(xì)胞系相關(guān)產(chǎn)品:Jurkat Clone E6-1細(xì)胞、KMS26細(xì)胞、NCIH1694細(xì)胞 NCI-H187細(xì)胞系相關(guān)產(chǎn)品:RCK8細(xì)胞、Y 1細(xì)胞、HEK-293H細(xì)胞 WISH細(xì)胞系相關(guān)產(chǎn)品:OLN 93細(xì)胞、OVCA433細(xì)胞、SW-1116細(xì)胞 MDA-MB-175-VII細(xì)胞系相關(guān)產(chǎn)品:SW-1463細(xì)胞、KOSC2細(xì)胞、NB.4細(xì)胞 NCI-H1155細(xì)胞系相關(guān)產(chǎn)品:H-146細(xì)胞、MDCK-II細(xì)胞、RSMC細(xì)胞 NCIH2227細(xì)胞系相關(guān)產(chǎn)品:OK細(xì)胞、RCC-JF細(xì)胞、HMrSV5細(xì)胞 Adeno-293細(xì)胞系相關(guān)產(chǎn)品:VMRC-LCD細(xì)胞、TSCC1細(xì)胞、8402細(xì)胞 Hs940-T細(xì)胞系相關(guān)產(chǎn)品:NCI-H2228細(xì)胞、SNU-886細(xì)胞、DMS 79細(xì)胞 UMR106細(xì)胞系相關(guān)產(chǎn)品:TGW-nu細(xì)胞、Panc203細(xì)胞、Mouse INsulinoma 6細(xì)胞 266 Bl細(xì)胞系相關(guān)產(chǎn)品:GM03671C細(xì)胞、KM-H2細(xì)胞、SACC-83細(xì)胞 Sol8 epithelioid cells小鼠骨骼肌肌肉母細(xì)胞系 KNS62細(xì)胞系相關(guān)產(chǎn)品:LLC-MK-2細(xì)胞、AML12細(xì)胞、SKNEP-1細(xì)胞 RAW264細(xì)胞系相關(guān)產(chǎn)品:TYK-nu細(xì)胞、CD18細(xì)胞、HIEC6細(xì)胞 PANC0813細(xì)胞系相關(guān)產(chǎn)品:293c18細(xì)胞、P3/ag細(xì)胞、BRL-3A細(xì)胞 H295R-S1細(xì)胞系相關(guān)產(chǎn)品:AU565細(xì)胞、AN3-CA細(xì)胞、HEK-2細(xì)胞 UMR106細(xì)胞系相關(guān)產(chǎn)品:TGW-nu細(xì)胞、Panc203細(xì)胞、Mouse INsulinoma 6細(xì)胞 EM-3細(xì)胞系相關(guān)產(chǎn)品:WERI-Rb-1細(xì)胞、SUP T1細(xì)胞、CNE-2細(xì)胞 W256細(xì)胞系相關(guān)產(chǎn)品:HUC細(xì)胞、SNU-739細(xì)胞、SKMES1細(xì)胞 NCM460細(xì)胞系相關(guān)產(chǎn)品:Metastatic Variant 522細(xì)胞、Panc-02細(xì)胞、NCI-H774細(xì)胞 MC38細(xì)胞系相關(guān)產(chǎn)品:QG56細(xì)胞、H-1238細(xì)胞、OCM1A細(xì)胞 JTC-28細(xì)胞系相關(guān)產(chǎn)品:G 292 Clone A 141B1細(xì)胞、HSC2細(xì)胞、MD Anderson-Metastatic Breast-415細(xì)胞 HBL-1 [Human diffuse large B-cell lymphoma]細(xì)胞系相關(guān)產(chǎn)品:RPMI8402細(xì)胞、NIH3T3細(xì)胞、C 643細(xì)胞 UACC-812細(xì)胞系相關(guān)產(chǎn)品:HT144-mel細(xì)胞、CL 1-5細(xì)胞、H69C細(xì)胞 GT39細(xì)胞系相關(guān)產(chǎn)品:NCI.H460細(xì)胞、RPMI8226細(xì)胞、HCC1588細(xì)胞 Loucy細(xì)胞系相關(guān)產(chǎn)品:Hep G2/C3A細(xì)胞、Mc Ardle 7777細(xì)胞、Hs294T細(xì)胞 H676細(xì)胞系相關(guān)產(chǎn)品:HCC38細(xì)胞、SKMES-1細(xì)胞、BC-025細(xì)胞 Panc-8_13細(xì)胞系相關(guān)產(chǎn)品:HUV-EC-C細(xì)胞、KMST6細(xì)胞、NCIH838細(xì)胞 SUNE 1細(xì)胞系相關(guān)產(chǎn)品:Buffalo Rat Liver-3A細(xì)胞、SUDHL6細(xì)胞、GLAG-66細(xì)胞 NCI-H889細(xì)胞系相關(guān)產(chǎn)品:P3 88 D1細(xì)胞、MCF.10A細(xì)胞、BCAP37細(xì)胞 NCI-H711細(xì)胞系相關(guān)產(chǎn)品:P3/X63/Ag8細(xì)胞、BEL-7405細(xì)胞、EoL-1-cell細(xì)胞 GC1-SPG細(xì)胞系相關(guān)產(chǎn)品:SaOS-2細(xì)胞、SUM149-PT細(xì)胞、H-1048細(xì)胞 Panc327細(xì)胞系相關(guān)產(chǎn)品:KPL-1細(xì)胞、BEL-7402細(xì)胞、COLO-320HSR細(xì)胞 Rca-B細(xì)胞系相關(guān)產(chǎn)品:GT39細(xì)胞、MDA-MB 231細(xì)胞、NCI-H1792細(xì)胞 MDA-415細(xì)胞系相關(guān)產(chǎn)品:HUVEC細(xì)胞、Mouse INsulinoma 6細(xì)胞、C22 (Clara)細(xì)胞 FDCP1細(xì)胞系相關(guān)產(chǎn)品:RT4P細(xì)胞、CCRF/CEM細(xì)胞、COR-L23細(xì)胞 HRIF細(xì)胞系相關(guān)產(chǎn)品:CV-1細(xì)胞、Madin-Darby Bovine Kidney細(xì)胞、MCF-10A細(xì)胞 CAL 85-1細(xì)胞系相關(guān)產(chǎn)品:Intestinal Epithelioid Cell line No. 18細(xì)胞、HIBEpiC細(xì)胞、SCC-7細(xì)胞 HOSEpiC細(xì)胞系相關(guān)產(chǎn)品:Toledo細(xì)胞、Hs-27細(xì)胞、P-36細(xì)胞 EA hy 926細(xì)胞系相關(guān)產(chǎn)品:GM00637B細(xì)胞、SUDHL-6細(xì)胞、HS-68細(xì)胞 LP-1細(xì)胞系相關(guān)產(chǎn)品:KYSE520細(xì)胞、Colon26細(xì)胞、HIBEC細(xì)胞 OE21細(xì)胞系相關(guān)產(chǎn)品:NCI-H508細(xì)胞、ECV304細(xì)胞、H64細(xì)胞 Tissue Culture-1細(xì)胞系相關(guān)產(chǎn)品:CA-46細(xì)胞、HCC1419細(xì)胞、WC00059細(xì)胞 NEC8細(xì)胞系相關(guān)產(chǎn)品:EB3 [Human Burkitt lymphoma]細(xì)胞、Rca-B細(xì)胞、KYSE410細(xì)胞 COLO394細(xì)胞系相關(guān)產(chǎn)品:373 MG細(xì)胞、BE2-M17細(xì)胞、RGM1細(xì)胞 NTera2 cl.D1細(xì)胞系相關(guān)產(chǎn)品:PaTu8988s細(xì)胞、253J B-V細(xì)胞、CoC1細(xì)胞 L6細(xì)胞系相關(guān)產(chǎn)品:SKCO 1細(xì)胞、DU4475細(xì)胞、Michigan Cancer Foundation-12F細(xì)胞 A 253細(xì)胞系相關(guān)產(chǎn)品:P31 FUJ細(xì)胞、HFLS-RA細(xì)胞、FOXNY細(xì)胞 PNEC30細(xì)胞系相關(guān)產(chǎn)品:Ly7細(xì)胞、FHC細(xì)胞、BCPAP細(xì)胞 HPC-Y5細(xì)胞系相關(guān)產(chǎn)品:GM02131A細(xì)胞、H1694細(xì)胞、IEC 6細(xì)胞 RPMI 1846細(xì)胞系相關(guān)產(chǎn)品:AMC-HN8細(xì)胞、CMT64細(xì)胞、TE85細(xì)胞 PC3M細(xì)胞系相關(guān)產(chǎn)品:BMSC/hBMSCs細(xì)胞、KM12SM細(xì)胞、PNT-1A細(xì)胞 Panc 03.27細(xì)胞系相關(guān)產(chǎn)品:Scott細(xì)胞、PANC-04-03細(xì)胞、MAVER1細(xì)胞 AML-EOL-1細(xì)胞系相關(guān)產(chǎn)品:HSAS1細(xì)胞、FAK+/+細(xì)胞、NCI-H2009細(xì)胞 Ishikawa細(xì)胞系相關(guān)產(chǎn)品:C166細(xì)胞、CCD19Lu細(xì)胞、SUM-102PT細(xì)胞 Hs 636 T細(xì)胞系相關(guān)產(chǎn)品:SNG-M細(xì)胞、Hs.27細(xì)胞、A-101D細(xì)胞 NCI-H2108細(xì)胞系相關(guān)產(chǎn)品:COLO 684細(xì)胞、D6P2T細(xì)胞、University of Arizona Cell Culture-893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 NTERA-2 clone D1細(xì)胞系相關(guān)產(chǎn)品:NCI-H226細(xì)胞、H-226細(xì)胞、C81-61細(xì)胞 PLCPRF5細(xì)胞系相關(guān)產(chǎn)品:SCL-II細(xì)胞、QBC939細(xì)胞、L929(NCTC)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 ACCM細(xì)胞系相關(guān)產(chǎn)品:Rat Skin 1細(xì)胞、143B細(xì)胞、SW626細(xì)胞 PANC1005細(xì)胞系相關(guān)產(chǎn)品:H-2198細(xì)胞、N-87細(xì)胞、MOLM-16細(xì)胞 Sol8 epithelioid cells小鼠骨骼肌肌肉母細(xì)胞系 BC-022細(xì)胞系相關(guān)產(chǎn)品:EL-4細(xì)胞、COLO 206F細(xì)胞、RPMI-7666細(xì)胞 KYSE 70細(xì)胞系相關(guān)產(chǎn)品:REC細(xì)胞、MFC細(xì)胞、NCI-SNU-C2B細(xì)胞 N1S1細(xì)胞系相關(guān)產(chǎn)品:NTera2 cl.D1細(xì)胞、CEMC7細(xì)胞、OV2008細(xì)胞 WISH細(xì)胞系相關(guān)產(chǎn)品:OLN 93細(xì)胞、HEK293EBNA細(xì)胞、HNE3細(xì)胞 "
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