2014年-2015工作文件优宁维学术-技术总介绍

CellseparationwithMACS®公司创始 美天旎全球总部位于德国科隆..我们的全球雇员：1200人，30%致力研发，分支机构遍布18个国2000多种产品销往全球40多个国家和地公我们的细治疗平台 公司 专利技细胞分选的金标准——MACS®无论手动分选，自动分选，均可高速，高纯度到目公司 技MACS®技术的组成部分：微珠、分选器和分选公司 产临床级细胞分选的GentleMACSTM全自动组织处理MACSQuantTM流式细细胞培养、扩增、刺激和公司的专利产美天旎中 美天旎中国目前有近5，售品，市场与技术服务， 。 优宁维生物科美天旎产品解决 Miltenyi提供一整套完整解决方案美天旎成长策 MACS®细胞分选的金标准MACS®技术原MACS®MagneticActivatedCell免疫学+细胞生物学+MACS®技术原MACS®MACS® MACSMACS

在高强度、梯度细MACS®直径50nm，对细胞无毒，可生MACS微直标多选

微珠通过偶联磁性标记MACS间标微抗FITC微 抗APC微 抗PE微亲和素微 抗生物素微 抗免疫球蛋白微MACS®间标磁珠：极大地扩展了分选在没有直接标记微珠使用自备抗体或配体的磁性分选分选或去除用多个抗体混合物标记的多分选稀有细胞和抗原弱表达的细MACS分选填充有磁性铁珠，有亲包被，对细胞MACS分选能进行完全 MACS分选常用MACS分选类最大细胞上样

最大细胞容

分子生物学应用MACSμ和M 10μgmRNA 50μgmRNA如何选择适合的MACS®分选分选柱容

ColumnDecision阳性

用于阳性分选的MACS®分选MACS®技术富集稀有细CD133+细胞(外周血DC细胞(外周血抗原特异T磁性标

磁性分

MACS®分选MACS®分选进行多参数分选的MACS®CellIsolationKitsCD4+CD25+RegulatoryTCellIsolationKit,CD4+CD25+RegulatoryTCellIsolationKit,DiamondPlasmacytoidDCIsolationKit,→AlwaysfollowDataMS,LS,XSLD,CS,D最常使用的分选柱类型MSandMACS®LargeCell分选柱内基质的孔径分选大细megakaryocyteswithCD61LangerhanscellsfromskinwithCD1aautoMACS™forupto4x109totalmax.2x108retained选择适合的autoMACS程accordingtoMACSreagentdataautoMACSUserManual,decisiontreeWholeBloodColumnKit(#130-093-试剂盒10WholeBlood50mLWholeBloodColumnElution适配的MACS®分选器MidiMACSQuadroMACS™分选VarioMACS™incombinationwithLSColumnSuperMACSII分选incombinationwithAdaperforMS,LSandLDWholeBloodColumnKit(#130-093-特殊的两层基质，优化全血样本的流ABMACS®MiniMACS™和OctoMACS™分选MS分选柱，LargeCell分选柱，MMidiMACS™和MACS®分选autoMACSTMautoMACSTMPro自动分选CliniMACSTMPlus分选thermoMACSTMMultiMACSTMMACS与FACS比MACS分选与FACS均以抗原-抗体反应为基均为高纯度流式细胞仪FACS

FACSFACS分选系FluidicDroplet可同时进行双项-4分项体小项体小，台式小，台式小，台式大激光类低功气冷低功率气冷低功气冷高功冷光固固固需要调激光488nm488nm数据处理式模数数模脱机否是是否最多检测487分选快》FACS分选时间（FACS每秒钟获取70000108cells=>23109cells=>4（没有包括操作人员调机器时间

》MACS®分选时间:(autoMACS™分选仪)平均每秒钟处理107细胞108cells=>5109cells=>5（无需调节机器操作简单、无需专人操作，可随时使无菌分选：仪器小巧毒方便，耗材为无菌性包MACS优势中尤为明显获取0分选与分析机器分开，节 、人力和时成分可以继续做磁性标记低成本（抗体108/支,磁珠109/支可以与流式分选联合使用，处理大量、低细胞MACSFACS分选三个参数以下分选性标志，仅需个参数分选高纯度、高回CD34+细胞的Original CD34StemCells parisonofPurityandEnri entofCD34+CellsfromBoneMarrow,UmbilicalCordandPeripheralBlood(PrimedforAphresis)UsingFiveSeparationSystems》比较了包括FACS与MACS在内的五种方法分选不依赖可 分选标志的特异分选标志表达很弱（如调节性T细胞低频细胞：CD133分选与应 标志在其它细胞上也有表MACS®技术分选人类DC分选单核细的

阳性分外周血分选DC细胞亚CD25分选 CD4+CD25+细细胞活性好，功能保留J.Immunol.199715938233837《CalciumIonophore-TreatedPeripheralBloodMonocytesandDendriticCellsRapidlyDisplayCharacteristicsofActivatedDendriticCells.》一文中，作者发现，Am. . ato196,10:21135pessonPaten and Celluar On f tokines n theNomaladToopsmaondnfetedmneban》一文中，作者最初想使用FACS分选小鼠大脑实质中分选细胞，但是没有活性细胞，可能因为分选时间长细胞在管道中受压力大，造成粒细胞凋亡。后改用温和AS。适用范 从到临床：CliniMACS是目前唯一一种获 面世以来，MiltenyiBiotec的各类 MACS产品介1、细 产2、细胞分选3、细胞分析4、分子生物学5、细胞培养1、MACS®样 产MACS®样 gentleMACS MACS®分选滤MACS®样 DeadCellRemovalRedBloodCellLysisBasicCD15MACSiBead™EndotoxinRemovalFcRBlockingReagentFcRBlockingReagent2、细胞分选人类细胞分选小鼠细胞分选大鼠细胞分选灵长类分选间接标记干祖细胞分干祖细胞内皮祖细胞分选间充质干细胞分选造血干祖细胞分使用CD34,CD133,CD117或MACS在干细胞研究中CD34+细胞的Original CD34StemCells parisonofPurityandEnri entofCD34+CellsfromBoneMarrow,UmbilicalCordandPeripheralBlood(PrimedforAphresis)UsingFive CD133（专利产品）：干细胞分骨脐带血、周CD34bright前（CD34brhtCndH-Rneg/dimCD90+和CD11+、 干胞皮胞2个不同的抗原表位：CD133/1（AC133）和CD133/2（293C3和MACS在MSC研究中anti-Fibroblast（培养后表型：CD34–,CD133–,CD14–,CD45–,CD105(SH2)+,CD166+,CD73(SH3uSH4)+,CD44+,CD90+2、来源：骨髓、脐带3、分类：非同源的多能干细胞，多种名4、作用：体外支持HSC扩增、体内促进HSC植活、组织再生、释放疗分子、免疫5、应用科研体外培养分化为多种中胚层的成熟细胞：神经细胞、肝细胞、心细胞、骨细胞、软骨细胞、脂肪细胞植入NOD/SCID小鼠及其它动物模型进行体内分化研临CCD3CD13MSCResearchToolBox-CD271FcRFcRNHExpansionMedium(2x500CytoMix-CD271NHOsteoDiffMedium(12x5Voucher(10%)foroneorderoftheVoucher(10%)foroneorderoftheµMACSOne-stepcDNAStarterPIQORStemCellMicroarrayKit,humanLNGFR-LNGFR-LNGFR-MSCResearchToolBox–CD271(LNGFR) LNGFR-LNGFR-LNGFR-CD45- CD45- CD45-Purity:85%(Ø83%±4%,n=4).EvaluationofseparationofCD271+cells–(CFU-FPlasticCFU-Fcountafter14daysofincubationandrelatedtothetotalnumberof1x106cells.ThenumbersofCFU-Frisesbyabout2logwhenBMMNCswereenrichedwithCD271(LNGFR)-APCandAnti-APC-MicroBeadsMACS小鼠干细胞分选试剂LineageCellDepletionKit:CD117MicroBeads：分选早期Anti-Sca-1MicroBeads：分选造血干细胞和早期造

小鼠骨髓造血干细胞(HSC)定Sca-1CD117+(c-kit)、Thy-1low和lin-细胞. T细胞分选产MACS®技术分选与分析功能性T

T细胞研究调节性T细胞的分选和功CD4+CD25+RegulatoryTCellIsolationCD4+CD25+CD127dim/-RegulatoryTCellIsolation新：CD4+CD25+CD45RA+RegulatoryTCellIsolationKit,人Treg分析的抗体和试-如Anti-FoxP3CD4CD25TregBefore IsolatedregulatoryTKit,人富集 去除非CD4+和CD127high细Tregs和效应性T细胞(e.g.CD4+T细胞,或CD4+CD25-T细胞在多克隆刺激物作用下，以一方案TregSuppressionInspectorTregSuppressionTregSuppression

TregSuppression

TregSuppression + 用TregSuppression检测分选后的调节性T细胞CD4+CD25+调节性T细胞试剂RatioRatioTrespvs.TregTregSuppression用于体外检测人CD4CD25+调节性TCD2,CD3andCD28抗体负载到抗BiotinMACSiBead™颗粒上。T细胞研究的创新性分选和分析抗原特异性T用Anti-IFN--PE进行胞内细胞 stimulated分选和分析抗原特异性T经过PepTivator多肽库刺激后的细胞分析和分选抗原CD4+T细胞CD137为基础，分选和分析活化的，抗原特异性CD8+T细胞因子分泌分选和分析细胞因子CytokineSecretionAssay–灵敏度高MACS细胞因子分泌实验•

MACS®细胞因子分泌细胞分析与分分选与检测试IL-2分泌IL-4分泌IL-5分泌IL-10分泌细IL-12分泌细IL-13分泌细TNF-分泌细

选与检测试IL-2分泌细IL-5分泌细IL-17分泌细胞B细胞分B细胞B细胞浆细B细胞发BCellIsolationKitCD19MicroBeads,Positiveselection 130-BCellIsolationKitCD19MicroBeads,Positiveselection 130-depletionofhumanB OrderWholeBloodCD19MicroBeads,IsolationofCD19+ 130-fromhumanwhole andbonemarrowusingtheautoMACS™ProSeparatororautoMACSSeparatorOrderCD19MultiSortKit,IsolationofhumanB 130- OrderCD20MicroBeads,Positiveselection 130-depletionofhumanB OrderCD22MicroBeads,Positiveselection 130-depletionofhumanB OrderBCellIsolationKitII,Isolationof 130-humanBcellsfrom bydepletionofnon-BOrderCD19MicroBeads,PositiveselectionordepletionofhumanBcellsOrder130-OrderWholeBloodCD19IsolationofCD19+130-fromhumanwholeandbonemarrowusingautoMACS™ProorautoMACSOrderCD19MultiSortKit,IsolationofhumanBcell130-OrderCD20MicroBeads,PositiveselectionordepletionofhumanBcells130-OrderCD22MicroBeads,PositiveselectionordepletionofhumanBcells130-BCellIsolationKitOrderBCellIsolationKitII,Isolationof130-humanBcellsfrombydepletionofnon-BCD69MicroBeadBCD69MicroBeadCD69MicroBeadCD27CD69MicroBeadKit,CD27CD69MicroBeadKit,CD27OrderCD27MicroBeads,Positiveselection130-depletionofmemoryBCD30OrderCD30MicroBeads,Positiveselection130-depletionofasubsetactivatedhumanBCD38MicroBeadOrderCD38MicroBeadKit,Positiveselection130-depletionofasubsetofhumanBCD43OrderCD43MicroBeads,Isolationof130-restinghumanBcellsdepletionofCD43+CD27MicroBeads,PositiveselectionCD27MicroBeads,Positiveselection 130-depletionof memoryBOrderAnti-IgGCD30CD30CD30MicroBeads,PositiveselectionCD30MicroBeads,Positiveselection 130-depletionofasubset activatedhumanBOrderAnti-IgMCD38MicroBeadAnti-IgMCD38MicroBeadAnti-IgMMicroBeads,OrderNaiveBCellIsolationKitOrderNaiveBCellIsolationKitCD38MicroBeadCD38MicroBeadKit,Positiveselection 130-depletionofa subsetofhumanBCD43NaiveBCellIsolationKitII,CD43NaiveBCellIsolationKitII,CD43MicroBeads,Isolationof 130-restingCD43MicroBeads,Isolationof 130-restinghumanBcells depletionofCD43+OrderMemoryBCellIsolationMemoryBCellIsolationKit,MemoryMemoryBCellIsolationKit,

PositiveselectionordepletionofasubsetofactivatedBcellsPositiveselectionordepletionPositiveselectionordepletionofasubsetofactivatedBcellsPositiveselectionordepletionofIgG+memoryBcellsPositiveselectionordepletionPositiveselectionordepletionofIgG+memoryBcellsPositiveselectionordepletionofIgM+cellsPositiveselectionordepletionPositiveselectionordepletionofIgM+cellsIsolationofuntouchednaiveBcellsfromPBMCsbydepletionofnon-BcellsandCD27+Bcells.IsolationofuntouchednaiveBcellsfromPBMCsbyIsolationofuntouchednaiveBcellsfromPBMCsbydepletionofnon-BcellsandCD27+Bcells.IsolationofhumanIsolationofhumanmemoryIsolationofhuman

Order130-Order130-Order130-Order130-OrderOrderOrderOrder130-130-OrderOrderOrder130-130-130-Order130-OrderCD27CD38MicroBeadCD138WholeBloodCD138PlasmaCellIsolationCD27MicroBeads,Positiveselection 130-depletionofCD27CD38MicroBeadCD138WholeBloodCD138PlasmaCellIsolationCD27MicroBeads,Positiveselection 130-depletionof memoryBOrderCD38MicroBeadKit,Positiveselection 130-depletionofa subsetofhumanBOrder130-CD138MicroBeads+CD138-PE,IsolationandfluorescentstainingofhumanplasmaCD138MicroBeads,Positiveselection 130-depletionofhuman OrderWholeBloodCD138MicroBeads,IsolationofCD138+ 130-fromhumanwhole andbonemarrowusingtheautoMACS™ProSeparatororautoMACSSeparatorOrderPlasmaCellIsolationKit,Isolationofhuman 130-plasma OrderCD27OrderCD27MicroBeads,Positiveselection130-depletionofmemoryBCD38MicroBeadOrderCD38MicroBeadKit,Positiveselection130-depletionofasubsetofhumanBCD138OrderCD138MicroBeads,Positiveselection130-depletionofhumanCD138MicroBeads+CD138-Isolationand130-stainingofhumanWholeBloodCD138OrderWholeBloodCD138IsolationofCD138+130-fromhumanwholeandbonemarrowusingautoMACS™ProorautoMACSPlasmaCellIsolationPlasmaCellIsolationKit,IsolationofhumanCD38+plasmacellsOrder130-DC细胞分析与分选识别人类DC和DCDC亚群特异性标志BloodDendriticLineage-(CD3,CD14,CD16,CD19,CD20,

BDCA-

BDCA-3(CD141)+BDCA-3(CD141)+分选人类DC和DCDC亚群特异性标志BloodDendriticLineage-(CD3,CD14,CD16,CD19,CD20,CD56)HLA-

CD303(BDCA-2)+CD304(BDCA-4)+PDCIsolationKitDiamondPDCIsolation

CD1c(BDCA-Isolation

MBAveragefrequenciesofDCsubsetsinfreshbloodandPBMCDC±0.140.41±0.17±0.160.62±0.01±0.020.04±0.01MACS®技术分选与分析人类DCCD14,CD19, CD14,CD19,DMDC-MDC-2MACS®技术分选人类DC分选前单核细胞的

阳性分外周血分选后DC细胞亚MACS®技术分选人类DCBDCA-1选MDC-

BDCA-3MDC-BDCA-4选CD8+CD4+mPDCA-CD8+CD4+mPDCA-CD11c+PlasmacytoidDCPlasmacytoidDCllmidDC)mloidDC)CD11cCD11c应用CD11c微珠阳性分选小鼠 MACS®技术分选与分析小鼠DCMACS®技术分选与分析小鼠pDC MACS®技术分选与分析大鼠DCAnti-DC(OX62单克隆抗体OX62识别表达于大部分大鼠DC上的整素αE2亚单位（分子量150kDa）两个主要的树突状细胞亚群：CD4/OX41双阳性CD4/OX41 体前叶星状滤泡样（S100+）细胞的一个亚群，但在MACS