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- 详细信息
- 技术资料
- 细胞类型:
细胞系
- 供应商:
智立中特(武汉)生物科技有限公司
- 库存:
100
- 英文名:
OCI-Ly3
- 运输方式:
顺丰派送
- 器官来源:
人
- 物种来源:
人
- 组织来源:
血液、淋巴系统
- 规格:
1×10⁶cells/T25培养瓶
细胞名称
人弥漫大B淋巴瘤细胞OCI-Ly3
产品规格
T25培养瓶x1
1.5ml冻存管x2
细胞数量
1x10^6;1x10^6
保存温度
37℃;-198℃
运输方式
常温保温运输;干冰运输
安全等级1
用途限制
仅供科研用途2类
培养体系
DMEM高糖培养基+20%FBS+1%三抗
培养温度 37℃
二氧化碳浓度 5%
简介
人弥漫大B淋巴瘤细胞OCI-Ly3细胞于1983年从52岁男性B细胞非霍奇金淋巴瘤骨髓样本中建立。为常规的药物筛选模型,悬浮培养,易聚团。引种自DSMZ(ACC-761)
注释
Partof:CancerCellLineEncyclopedia(CCLE)project.
Partof:LL-100bloodcancercelllinepanel.
Partof:MDAndersonCellLinesProject.
From:OntarioCancerInstitute(OCI);Toronto;Canada.
Doublingtime:~24hours(DSMZ).
Omics:Array-basedCGH.
Omics:CNVanalysis.
Omics:Deepexomeanalysis.
Omics:DeepRNAseqanalysis.
Omics:Genomesequenced.
Omics:H3K9acChIP-seqepigenomeanalysis.
Omics:miRNAexpressionprofiling.
Omics:Proteinexpressionbyreverse-phaseproteinarrays.
Omics:SNParrayanalysis.
Omics:Transcriptomeanalysis.
Caution:Acellline,nowtermedGNE-587170(CVCL_AT69),wasmistakenlydistributedbyOCItoanumberofgroupsunderthedesignationOCI-Ly-3,theoriginofthatcelllineisnowknown.
Derivedfromsamplingsite:Bonemarrow.
STR信息
AmelogeninX,Y
CSF1PO10,12
D5S81812
D7S82011,12
D13S31711
D16S53911,12
TH017,9.3
TPOX8,12
vWA17
参考文献
PubMed=3567358
TweeddaleM.E.,LimB.,JamalN.,RobinsonJ.,ZalcbergJ.R.,LockwoodG.,MindenM.D.,MessnerH.A.
Thepresenceofclonogeniccellsinhigh-grademalignantlymphoma:aprognosticfactor.
Blood69:1307-1314(1987)
PubMed=8574164;DOI=10.3109/10428199509059672
ChangH.,BlondalJ.A.,BenchimolS.,MindenM.D.,MessnerH.A.
p53mutations,c-mycandbcl-2rearrangementsinhumannon-Hodgkin'slymphomacelllines.
Leuk.Lymphoma19:165-171(1995)
PubMed=10676951;DOI=10.1038/35000501
AlizadehA.A.,EisenM.B.,DavisR.E.,MaC.,LossosI.S.,RosenwaldA.,BoldrickJ.C.,SabetH.,TranT.,YuX.,PowellJ.I.,YangL.,MartiG.E.,MooreT.,HudsonJ.Jr.,LuL.,LewisD.B.,TibshiraniR.,SherlockG.,ChanW.C.,GreinerT.C.,WeisenburgerD.D.,ArmitageJ.O.,WarnkeR.,LevyR.,WilsonW.,GreverM.R.,ByrdJ.C.,BotsteinD.,BrownP.O.,StaudtL.M.
DistincttypesofdiffuselargeB-celllymphomaidentifiedbygeneexpressionprofiling.
Nature403:503-511(2000)
PubMed=11807979;DOI=10.1002/gcc.10025
MehraS.,MessnerH.A.,MindenM.D.,ChagantiR.S.K.
Molecularcytogeneticcharacterizationofnon-Hodgkinlymphomacelllines.
GenesChromosomesCancer33:225-234(2002)
PubMed=19278952;DOI=10.1182ood-2009-01-202028
LiC.,KimS.-W.,RaiD.,BollaA.R.,AdhvaryuS.,KinneyM.C.,RobetoryeR.S.,AguiarR.C.T.
Copynumberabnormalities,MYCactivity,andthegeneticfingerprintofnormalBcellsmechanisticallydefinethemicroRNAprofileofdiffuselargeB-celllymphoma.
Blood113:6681-6690(2009)
PubMed=22460905;DOI=10.1038/nature11003
BarretinaJ.G.,CaponigroG.,StranskyN.,VenkatesanK.,MargolinA.A.,KimS.,WilsonC.J.,LeharJ.,KryukovG.V.,SonkinD.,ReddyA.,LiuM.,MurrayL.,BergerM.F.,MonahanJ.E.,MoraisP.,MeltzerJ.,KorejwaA.,Jane-ValbuenaJ.,MapaF.A.,ThibaultJ.,Bric-FurlongE.,RamanP.,ShipwayA.,EngelsI.H.,ChengJ.,YuG.K.,YuJ.,AspesiP.Jr.,deSilvaM.,JagtapK.,JonesM.D.,WangL.,HattonC.,PalescandoloE.,GuptaS.,MahanS.,SougnezC.,OnofrioR.C.,LiefeldT.,MacConaillL.E.,WincklerW.,ReichM.,LiN.,MesirovJ.P.,GabrielS.B.,GetzG.,ArdlieK.,ChanV.,MyerV.E.,WeberB.L.,PorterJ.,WarmuthM.,FinanP.,HarrisJ.L.,MeyersonM.,GolubT.R.,MorrisseyM.P.,SellersW.R.,SchlegelR.,GarrawayL.A.
TheCancerCellLineEncyclopediaenablespredictivemodellingofanticancerdrugsensitivity.
Nature483:603-607(2012)
PubMed=23292937;DOI=10.1073/pnas.1205299110
ZhangJ.,GruborV.,LoveC.L.,BanerjeeA.,RichardsK.L.,MieczkowskiP.A.,DunphyC.,ChoiW.,AuW.Y.,SrivastavaG.,LugarP.L.,RizzieriD.A.,LagooA.S.,Bernal-MizrachiL.,MannK.P.,FlowersC.,NareshK.,EvensA.,GordonL.I.,CzaderM.B.,GillJ.I.,HsiE.D.,LiuQ.,FanA.,WalshK.,JimaD.,SmithL.L.,JohnsonA.J.,ByrdJ.C.,LuftigM.A.,NiT.,ZhuJ.,ChadburnA.,LevyS.,DunsonD.B.,DaveS.S.
GeneticheterogeneityofdiffuselargeB-celllymphoma.
Proc.Natl.Acad.Sci.U.S.A.110:1398-1403(2013)
PubMed=23699601;DOI=10.1182ood-2013-02-483727
MorinR.D.,MungallK.,PleasanceE.D.,MungallA.J.,GoyaR.,HuffR.D.,ScottD.W.,DingJ.,RothA.,ChiuR.,CorbettR.D.,ChanF.C.,Mendez-LagoM.,TrinhD.L.,Bolger-MunroM.,TaylorG.,HadjKhodabakhshiA.,Ben-NeriahS.,PonJ.,MeissnerB.,WoolcockB.,FarnoudN.,RogicS.,LimE.L.,JohnsonN.A.,ShahS.,JonesS.,SteidlC.,HoltR.,BirolI.,MooreR.,ConnorsJ.M.,GascoyneR.D.,MarraM.A.
MutationalandstructuralanalysisofdiffuselargeB-celllymphomausingwhole-genomesequencing.
Blood122:1256-1265(2013)
PubMed=25485619;DOI=10.1038/nbt.3080
KlijnC.,DurinckS.,StawiskiE.W.,HavertyP.M.,JiangZ.,LiuH.,DegenhardtJ.,MaybaO.,GnadF.,LiuJ.,PauG.,ReederJ.,CaoY.,MukhyalaK.,SelvarajS.K.,YuM.,ZyndaG.J.,BrauerM.J.,WuT.D.,GentlemanR.C.,ManningG.,YauchR.L.,BourgonR.,StokoeD.,ModrusanZ.,NeveR.M.,deSauvageF.J.,SettlemanJ.,SeshagiriS.,ZhangZ.
Acomprehensivetranscriptionalportraitofhumancancercelllines.
Nat.Biotechnol.33:306-312(2015)
PubMed=28196595;DOI=10.1016/j.ccell.2017.01.005
LiJ.,ZhaoW.,AkbaniR.,LiuW.,JuZ.,LingS.,VellanoC.P.,RoebuckP.,YuQ.,EterovicA.K.,ByersL.A.,DaviesM.A.,DengW.,GopalY.N.V.,ChenG.,vonEuwE.M.,SlamonD.J.,ConklinD.,HeymachJ.V.,GazdarA.F.,MinnaJ.D.,MyersJ.N.,LuY.,MillsG.B.,LiangH.
Characterizationofhumancancercelllinesbyreverse-phaseproteinarrays.
CancerCell31:225-239(2017)
PubMed=30285677;DOI=10.1186/s12885-018-4840-5
TanK.-T.,DingL.-W.,SunQ.-Y.,LaoZ.-T.,ChienW.,RenX.,XiaoJ.-F.,LohX.-Y.,XuL.,LillM.,MayakondaA.,LinD.-C.,YangH.,KoefflerH.P.
ProfilingtheB/Tcellreceptorrepertoireoflymphocytederivedcelllines.
BMCCancer18:940-940(2018)
PubMed=30894373;DOI=10.1158/0008-5472.CAN-18-2747
DutilJ.,ChenZ.,MonteiroA.N.,TeerJ.K.,EschrichS.A.
Aninteractiveresourcetoprobegeneticdiversityandestimatedancestryincancercelllines.
CancerRes.79:1263-1273(2019)
PubMed=31068700;DOI=10.1038/s41586-019-1186-3
GhandiM.,HuangF.W.,Jane-ValbuenaJ.,KryukovG.V.,LoC.C.,McDonaldE.R.III,BarretinaJ.,GelfandE.T.,BielskiC.M.,LiH.,HuK.,Andreev-DrakhlinA.Y.,KimJ.,HessJ.M.,HaasB.J.,AguetF.,WeirB.A.,RothbergM.V.,PaolellaB.R.,LawrenceM.S.,AkbaniR.,LuY.,TivH.L.,GokhaleP.C.,deWeckA.,MansourA.A.,OhC.,ShihJ.,HadiK.,RosenY.,BistlineJ.,VenkatesanK.,ReddyA.,SonkinD.,LiuM.,LeharJ.,KornJ.M.,PorterD.A.,JonesM.D.,GoljiJ.,CaponigroG.,TaylorJ.E.,DunningC.M.,CreechA.L.,WarrenA.C.,McFarlandJ.M.,ZamanighomiM.,KauffmannA.,StranskyN.,ImielinskiM.,MaruvkaY.E.,CherniackA.D.,TsherniakA.,VazquezF.,JaffeJ.D.,LaneA.A.,WeinstockD.M.,JohannessenC.M.,MorrisseyM.P.,StegmeierF.,SchlegelR.,HahnW.C.,GetzG.,MillsG.B.,BoehmJ.S.,GolubT.R.,GarrawayL.A.,SellersW.R.
Next-generationcharacterizationoftheCancerCellLineEncyclopedia.
Nature569:503-508(2019)
PubMed=31160637;DOI=10.1038/s41598-019-44491-x
QuentmeierH.,PommerenkeC.,DirksW.G.,EberthS.,KoeppelM.,MacLeodR.A.F.,NagelS.,SteubeK.,UphoffC.C.,DrexlerH.G.
TheLL-100panel:100celllinesforbloodcancerstudies.
Sci.Rep.9:8218-8218(2019)
验收细胞注意事项
1、收到人弥漫大B淋巴瘤细胞OCI-Ly3细胞,请查看瓶子是否有破裂,培养基是否漏出,是否浑浊,如有请尽快联系。
2、收到人弥漫大B淋巴瘤细胞OCI-Ly3细胞,如包装完好,请在显微镜下观察细胞。,由于运输过程中的问题,细胞培养瓶中的贴壁细胞有可能从瓶壁中脱落下来,显微镜下观察会出现细胞悬浮的情况,出现此状态时,请不要打开细胞培养瓶,应立即将培养瓶置于细胞培养箱里静止3-5小时左右,让细胞先稳定下,再于显微镜下观察,此时多数细胞会重新贴附于瓶壁。如细胞仍不能贴壁,请用台盼蓝染色法鉴定细胞活力,如台盼蓝染色证实细胞活力正常请按悬浮细胞的方法处理。
3、收到人弥漫大B淋巴瘤细胞OCI-Ly3细胞后,请镜下观察细胞,用恰当方式处理细胞。若悬浮的细胞较多,请离心收集细胞,接种到一个新的培养瓶中。弃掉原液,使用新鲜配制的培养基,使用进口胎牛血清。刚接到细胞,若细胞不多时血清浓度可以加到15%去培养。若细胞迏到80%左右,血清浓度还是在10%。
4、收到人弥漫大B淋巴瘤细胞OCI-Ly3细胞时如无异常情况,请在显微镜下观察细胞密度,如为贴壁细胞,未超过80%汇合度时,将培养瓶中培养基吸出,留下5-10ML培养基继续培养:超过80%汇合度时,请按细胞培养条件传代培养。如为悬浮细胞,吸出培养液,1000转/分钟离心3分钟,吸出上清,管底细胞用新鲜培养基悬浮细胞后移回培养瓶。
5、将培养瓶置于37℃培养箱中培养,盖子微微拧松。吸出的培养基可以保存在灭菌过的瓶子里,存放于4℃冰箱,以备不时之需。
6、24小时后,细胞形态已恢复并贴满瓶壁,即可传代。(贴壁细胞)将培养瓶里的培养基倒去,加3-5ml(以能覆盖细胞生长面为准)PBS或Hanks’液洗涤后弃去。加0.5-1ml0.25%含EDTA的胰酶消化,消化时间以具体细胞为准,一般1-3分钟,不超过5分钟。可以放入37℃培养箱消化。轻轻晃动瓶壁,见细胞脱落下来,加入3-5ml培养基终止消化。用移液管轻轻吹打瓶壁上的细胞,使之完全脱落,然后将溶液吸入离心管内离心,1000rpm/5min。弃上清,视细胞数量决定分瓶数,一般一传二,如细胞量多可一传三,有些细胞不易传得过稀,有些生长较快的细胞则可以多传几瓶,以具体细胞和经验为准。(悬浮细胞)用移液管轻轻吹打瓶壁,直接将溶液吸入离心管离心即可。
7、贴壁细胞,悬浮细胞。严格无菌操作。换液时,换新的细胞培养瓶和换新鲜的培养液,37℃,5%CO2培养。
人弥漫大B淋巴瘤细胞OCI-Ly3
产品规格
T25培养瓶x1
1.5ml冻存管x2
细胞数量
1x10^6;1x10^6
保存温度
37℃;-198℃
运输方式
常温保温运输;干冰运输
安全等级1
用途限制
仅供科研用途2类
培养体系
DMEM高糖培养基+20%FBS+1%三抗
培养温度 37℃
二氧化碳浓度 5%
简介
人弥漫大B淋巴瘤细胞OCI-Ly3细胞于1983年从52岁男性B细胞非霍奇金淋巴瘤骨髓样本中建立。为常规的药物筛选模型,悬浮培养,易聚团。引种自DSMZ(ACC-761)
注释
Partof:CancerCellLineEncyclopedia(CCLE)project.
Partof:LL-100bloodcancercelllinepanel.
Partof:MDAndersonCellLinesProject.
From:OntarioCancerInstitute(OCI);Toronto;Canada.
Doublingtime:~24hours(DSMZ).
Omics:Array-basedCGH.
Omics:CNVanalysis.
Omics:Deepexomeanalysis.
Omics:DeepRNAseqanalysis.
Omics:Genomesequenced.
Omics:H3K9acChIP-seqepigenomeanalysis.
Omics:miRNAexpressionprofiling.
Omics:Proteinexpressionbyreverse-phaseproteinarrays.
Omics:SNParrayanalysis.
Omics:Transcriptomeanalysis.
Caution:Acellline,nowtermedGNE-587170(CVCL_AT69),wasmistakenlydistributedbyOCItoanumberofgroupsunderthedesignationOCI-Ly-3,theoriginofthatcelllineisnowknown.
Derivedfromsamplingsite:Bonemarrow.
STR信息
AmelogeninX,Y
CSF1PO10,12
D5S81812
D7S82011,12
D13S31711
D16S53911,12
TH017,9.3
TPOX8,12
vWA17
参考文献
PubMed=3567358
TweeddaleM.E.,LimB.,JamalN.,RobinsonJ.,ZalcbergJ.R.,LockwoodG.,MindenM.D.,MessnerH.A.
Thepresenceofclonogeniccellsinhigh-grademalignantlymphoma:aprognosticfactor.
Blood69:1307-1314(1987)
PubMed=8574164;DOI=10.3109/10428199509059672
ChangH.,BlondalJ.A.,BenchimolS.,MindenM.D.,MessnerH.A.
p53mutations,c-mycandbcl-2rearrangementsinhumannon-Hodgkin'slymphomacelllines.
Leuk.Lymphoma19:165-171(1995)
PubMed=10676951;DOI=10.1038/35000501
AlizadehA.A.,EisenM.B.,DavisR.E.,MaC.,LossosI.S.,RosenwaldA.,BoldrickJ.C.,SabetH.,TranT.,YuX.,PowellJ.I.,YangL.,MartiG.E.,MooreT.,HudsonJ.Jr.,LuL.,LewisD.B.,TibshiraniR.,SherlockG.,ChanW.C.,GreinerT.C.,WeisenburgerD.D.,ArmitageJ.O.,WarnkeR.,LevyR.,WilsonW.,GreverM.R.,ByrdJ.C.,BotsteinD.,BrownP.O.,StaudtL.M.
DistincttypesofdiffuselargeB-celllymphomaidentifiedbygeneexpressionprofiling.
Nature403:503-511(2000)
PubMed=11807979;DOI=10.1002/gcc.10025
MehraS.,MessnerH.A.,MindenM.D.,ChagantiR.S.K.
Molecularcytogeneticcharacterizationofnon-Hodgkinlymphomacelllines.
GenesChromosomesCancer33:225-234(2002)
PubMed=19278952;DOI=10.1182ood-2009-01-202028
LiC.,KimS.-W.,RaiD.,BollaA.R.,AdhvaryuS.,KinneyM.C.,RobetoryeR.S.,AguiarR.C.T.
Copynumberabnormalities,MYCactivity,andthegeneticfingerprintofnormalBcellsmechanisticallydefinethemicroRNAprofileofdiffuselargeB-celllymphoma.
Blood113:6681-6690(2009)
PubMed=22460905;DOI=10.1038/nature11003
BarretinaJ.G.,CaponigroG.,StranskyN.,VenkatesanK.,MargolinA.A.,KimS.,WilsonC.J.,LeharJ.,KryukovG.V.,SonkinD.,ReddyA.,LiuM.,MurrayL.,BergerM.F.,MonahanJ.E.,MoraisP.,MeltzerJ.,KorejwaA.,Jane-ValbuenaJ.,MapaF.A.,ThibaultJ.,Bric-FurlongE.,RamanP.,ShipwayA.,EngelsI.H.,ChengJ.,YuG.K.,YuJ.,AspesiP.Jr.,deSilvaM.,JagtapK.,JonesM.D.,WangL.,HattonC.,PalescandoloE.,GuptaS.,MahanS.,SougnezC.,OnofrioR.C.,LiefeldT.,MacConaillL.E.,WincklerW.,ReichM.,LiN.,MesirovJ.P.,GabrielS.B.,GetzG.,ArdlieK.,ChanV.,MyerV.E.,WeberB.L.,PorterJ.,WarmuthM.,FinanP.,HarrisJ.L.,MeyersonM.,GolubT.R.,MorrisseyM.P.,SellersW.R.,SchlegelR.,GarrawayL.A.
TheCancerCellLineEncyclopediaenablespredictivemodellingofanticancerdrugsensitivity.
Nature483:603-607(2012)
PubMed=23292937;DOI=10.1073/pnas.1205299110
ZhangJ.,GruborV.,LoveC.L.,BanerjeeA.,RichardsK.L.,MieczkowskiP.A.,DunphyC.,ChoiW.,AuW.Y.,SrivastavaG.,LugarP.L.,RizzieriD.A.,LagooA.S.,Bernal-MizrachiL.,MannK.P.,FlowersC.,NareshK.,EvensA.,GordonL.I.,CzaderM.B.,GillJ.I.,HsiE.D.,LiuQ.,FanA.,WalshK.,JimaD.,SmithL.L.,JohnsonA.J.,ByrdJ.C.,LuftigM.A.,NiT.,ZhuJ.,ChadburnA.,LevyS.,DunsonD.B.,DaveS.S.
GeneticheterogeneityofdiffuselargeB-celllymphoma.
Proc.Natl.Acad.Sci.U.S.A.110:1398-1403(2013)
PubMed=23699601;DOI=10.1182ood-2013-02-483727
MorinR.D.,MungallK.,PleasanceE.D.,MungallA.J.,GoyaR.,HuffR.D.,ScottD.W.,DingJ.,RothA.,ChiuR.,CorbettR.D.,ChanF.C.,Mendez-LagoM.,TrinhD.L.,Bolger-MunroM.,TaylorG.,HadjKhodabakhshiA.,Ben-NeriahS.,PonJ.,MeissnerB.,WoolcockB.,FarnoudN.,RogicS.,LimE.L.,JohnsonN.A.,ShahS.,JonesS.,SteidlC.,HoltR.,BirolI.,MooreR.,ConnorsJ.M.,GascoyneR.D.,MarraM.A.
MutationalandstructuralanalysisofdiffuselargeB-celllymphomausingwhole-genomesequencing.
Blood122:1256-1265(2013)
PubMed=25485619;DOI=10.1038/nbt.3080
KlijnC.,DurinckS.,StawiskiE.W.,HavertyP.M.,JiangZ.,LiuH.,DegenhardtJ.,MaybaO.,GnadF.,LiuJ.,PauG.,ReederJ.,CaoY.,MukhyalaK.,SelvarajS.K.,YuM.,ZyndaG.J.,BrauerM.J.,WuT.D.,GentlemanR.C.,ManningG.,YauchR.L.,BourgonR.,StokoeD.,ModrusanZ.,NeveR.M.,deSauvageF.J.,SettlemanJ.,SeshagiriS.,ZhangZ.
Acomprehensivetranscriptionalportraitofhumancancercelllines.
Nat.Biotechnol.33:306-312(2015)
PubMed=28196595;DOI=10.1016/j.ccell.2017.01.005
LiJ.,ZhaoW.,AkbaniR.,LiuW.,JuZ.,LingS.,VellanoC.P.,RoebuckP.,YuQ.,EterovicA.K.,ByersL.A.,DaviesM.A.,DengW.,GopalY.N.V.,ChenG.,vonEuwE.M.,SlamonD.J.,ConklinD.,HeymachJ.V.,GazdarA.F.,MinnaJ.D.,MyersJ.N.,LuY.,MillsG.B.,LiangH.
Characterizationofhumancancercelllinesbyreverse-phaseproteinarrays.
CancerCell31:225-239(2017)
PubMed=30285677;DOI=10.1186/s12885-018-4840-5
TanK.-T.,DingL.-W.,SunQ.-Y.,LaoZ.-T.,ChienW.,RenX.,XiaoJ.-F.,LohX.-Y.,XuL.,LillM.,MayakondaA.,LinD.-C.,YangH.,KoefflerH.P.
ProfilingtheB/Tcellreceptorrepertoireoflymphocytederivedcelllines.
BMCCancer18:940-940(2018)
PubMed=30894373;DOI=10.1158/0008-5472.CAN-18-2747
DutilJ.,ChenZ.,MonteiroA.N.,TeerJ.K.,EschrichS.A.
Aninteractiveresourcetoprobegeneticdiversityandestimatedancestryincancercelllines.
CancerRes.79:1263-1273(2019)
PubMed=31068700;DOI=10.1038/s41586-019-1186-3
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验收细胞注意事项
1、收到人弥漫大B淋巴瘤细胞OCI-Ly3细胞,请查看瓶子是否有破裂,培养基是否漏出,是否浑浊,如有请尽快联系。
2、收到人弥漫大B淋巴瘤细胞OCI-Ly3细胞,如包装完好,请在显微镜下观察细胞。,由于运输过程中的问题,细胞培养瓶中的贴壁细胞有可能从瓶壁中脱落下来,显微镜下观察会出现细胞悬浮的情况,出现此状态时,请不要打开细胞培养瓶,应立即将培养瓶置于细胞培养箱里静止3-5小时左右,让细胞先稳定下,再于显微镜下观察,此时多数细胞会重新贴附于瓶壁。如细胞仍不能贴壁,请用台盼蓝染色法鉴定细胞活力,如台盼蓝染色证实细胞活力正常请按悬浮细胞的方法处理。
3、收到人弥漫大B淋巴瘤细胞OCI-Ly3细胞后,请镜下观察细胞,用恰当方式处理细胞。若悬浮的细胞较多,请离心收集细胞,接种到一个新的培养瓶中。弃掉原液,使用新鲜配制的培养基,使用进口胎牛血清。刚接到细胞,若细胞不多时血清浓度可以加到15%去培养。若细胞迏到80%左右,血清浓度还是在10%。
4、收到人弥漫大B淋巴瘤细胞OCI-Ly3细胞时如无异常情况,请在显微镜下观察细胞密度,如为贴壁细胞,未超过80%汇合度时,将培养瓶中培养基吸出,留下5-10ML培养基继续培养:超过80%汇合度时,请按细胞培养条件传代培养。如为悬浮细胞,吸出培养液,1000转/分钟离心3分钟,吸出上清,管底细胞用新鲜培养基悬浮细胞后移回培养瓶。
5、将培养瓶置于37℃培养箱中培养,盖子微微拧松。吸出的培养基可以保存在灭菌过的瓶子里,存放于4℃冰箱,以备不时之需。
6、24小时后,细胞形态已恢复并贴满瓶壁,即可传代。(贴壁细胞)将培养瓶里的培养基倒去,加3-5ml(以能覆盖细胞生长面为准)PBS或Hanks’液洗涤后弃去。加0.5-1ml0.25%含EDTA的胰酶消化,消化时间以具体细胞为准,一般1-3分钟,不超过5分钟。可以放入37℃培养箱消化。轻轻晃动瓶壁,见细胞脱落下来,加入3-5ml培养基终止消化。用移液管轻轻吹打瓶壁上的细胞,使之完全脱落,然后将溶液吸入离心管内离心,1000rpm/5min。弃上清,视细胞数量决定分瓶数,一般一传二,如细胞量多可一传三,有些细胞不易传得过稀,有些生长较快的细胞则可以多传几瓶,以具体细胞和经验为准。(悬浮细胞)用移液管轻轻吹打瓶壁,直接将溶液吸入离心管离心即可。
7、贴壁细胞,悬浮细胞。严格无菌操作。换液时,换新的细胞培养瓶和换新鲜的培养液,37℃,5%CO2培养。
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人弥漫大B淋巴瘤细胞OCI-Ly3
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