Stem Cell quick guide 2013

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Mesenchymal stem cells Drivingyour MSC research Human samples Mouse samples Phenotyping of expanded MSCs Human • MSC Phenotyping Kit • Fluorochrome-conjugated antibodies against, e.g., CD73, CD90, CD105, CD45, CD34, CD14, CD20, etc. Mouse • Fluorochrome-conjugated antibodies against, e.g., Lineage markers, CD105, CD117, Anti-Sca-1, CD45, CD116, etc. • MACSQuant® Flow Cytometers Molecular analysis of MSCs • mRNA Isolation Kit and cDNA Synthesis Kit (for RT-PCR) • Protein Isolation Kits (for ChIP or normal IPs) • Microarray services • SuperAmp™ Technology • Bioinformatics • MultiMACS™ M96 Separator • Mitochondria isolation Differentiation of MSCs Human • NH Differentiation Media • Cytokines, e.g., TGF-β • NH CFU-F Mouse • Cytokines, e.g., FGF-8b Characterization of MSCs Sample type Frozen • Dead Cell Removal Kit • Pre-Separation Filters Tissues • gentleMACS™ Dissociator • Pre-Separation Filters Phenotyping of MSCs prior to expansion Fluorochrome-conjugated antibodies against, e.g., • CD271 • Anti-MSCA-1 • CD146 • CD105 • CD45 Phenotyping of MSCs prior to expansion Fluorochrome-conjugated antibodies against, e.g., • CD45 • Anti-Ter-119 • Anti-Sca-1 • CD105 Isolation of MSCs Magnetic isolation • CD271 (LNGFR) MicroBead Kits • Anti-MSCA-1 (W8B2) MicroBead Kit • CD90 MicroBeads • CD105 MicroBeads • CD146 MicroBead Kit • CD73 Antibodies • Lineage Cell Depletion Kit Flow sorting • Fluorochrome-conjugated antibodies (markers as listed) Plastic adherence • NH Expansion Media • CytoMix – MSC Expansion of MSCs • NH Expansion Media • CytoMix – MSC • Cytokines, e.g., FGF-2, PDGF-BB, TGF-β Isolation of MSCs Magnetic isolation • Lineage Cell Depletion Kit • CD105 MultiSort Kit (PE), CD117 MicroBeads, Anti-Sca-1 MicroBead Kit (FITC) • CD45 MicroBeads, Anti-Ter-119 MicroBeads removal pre-culture • CD11b MicroBeads removal post-culture Flow sorting • Fluorochrome- conjugated antibodies (markers as listed) Plastic adherence Expansion of MSCs • Cytokines, e.g., TGF-β, EGF Translate research to clinical applications • GMP Cytokines • CliniMACS® System • CliniMACS Reagents, e.g., CD271 Sample preparation Cell separation Cell analysis Cell culture Molecular applications Clinical applications Quick guide inside! MACS® Stem Cell Research macs-stemcells.com Germany/Austria/ Switzerland Miltenyi Biotec GmbH Friedrich-Ebert-Straße 68 51429 Bergisch Gladbach Germany Phone +49 2204 8306-0 Fax +49 2204 85197 macs@miltenyibiotec.de USA/Canada Miltenyi Biotec Inc. 2303 Lindbergh Street Auburn, CA 95602, USA Phone 800 FOR MACS Phone +1 530 888 8871 Fax +1 530 888 8925 macs@miltenyibiotec.com www.miltenyibiotec.com Australia Miltenyi Biotec Australia Pty. Ltd. Unit 16A, 2 Eden Park Drive North Ryde, NSW 2113, Australia Phone +61 2 8877 7400 Fax +61 2 9889 5044 macs@miltenyibiotec.com.au Benelux Miltenyi Biotec B.V. Schipholweg 68 H 2316 XE Leiden The Netherlands macs@miltenyibiotec.nl Customer service: The Netherlands Phone 0800 4020120 Fax 0800 4020100 Belgium Phone 0800 94016 Fax 0800 99626 Luxembourg Phone 800 24971 Fax 800 24984 China Miltenyi Biotec GmbH Shanghai Office Rm. 2309 No. 319 Xianxia Rd. Shanghai 200051, P.R. China Phone +86 21 62351005 Fax +86 21 62350953 macs@miltenyibiotec.com.cn France Miltenyi Biotec SAS 10 rue Mercoeur 75011 Paris, France Phone +33 1 56 98 16 16 Fax +33 1 56 98 16 17 macs@miltenyibiotec.fr Italy Miltenyi Biotec S.r.l. Via Persicetana, 2/D 40012 Calderara di Reno (BO) Italy Phone +39 051 6 460 411 Fax +39 051 6 460 499 macs@miltenyibiotec.it Japan Miltenyi Biotec K.K. Nittsu-Eitai Building 5F 16-10 Fuyuki, Koto-ku, Tokyo 135-0041, Japan Phone +81 3 5646 8910 Fax +81 3 5646 8911 macs@miltenyibiotec.jp Nordics and Baltics Miltenyi Biotec Norden AB Scheelevägen 17 223 70 Lund Sweden macs@miltenyibiotec.se Customer service: Sweden Phone 0200-111 800 Fax 046-280 72 99 Denmark Phone 80 20 30 10 Fax +46 46 280 72 99 Norway, Finland, Iceland, and Baltic countries Phone +46 46 280 72 80 Fax +46 46 280 72 99 Singapore Miltenyi Biotec Asia Pacific Pte Ltd. 100 Beach Road #28-06 to 28-08 Shaw Tower Singapore 189702 Phone +65 6238 8183 Fax +65 6238 0302 macs@miltenyibiotec.com.sg Spain Miltenyi Biotec S.L. C/Luis Buñuel 2 Ciudad de la Imagen 28223 Pozuelo de Alarcón (Madrid) Spain Phone +34 91 512 12 90 Fax +34 91 512 12 91 macs@miltenyibiotec.es United Kingdom Miltenyi Biotec Ltd. Almac House, Church Lane Bisley, Surrey GU24 9DR, UK Phone +44 1483 799 800 Fax +44 1483 799 811 macs@miltenyibiotec.co.uk 130-097-901.02 Miltenyi Biotec provides products and services worldwide. Visit www.miltenyibiotec.com/local to find your nearest Miltenyi Biotec contact. Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use. MACS, MACSelect, MACSQuant, CliniMACS, gentleMACS, MultiMACS, and SuperAmp are either registered trademarks or trademarks of Miltenyi Biotec GmbH. CryoStem, Pluriton, StainAlive, and Stemfect are all trademarks of Stemgent, Inc. Copyright © 2013 Miltenyi Biotec GmbH. All rights reserved.

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Sample preparation Cell separation Cell analysis Cell culture Molecular applications Clinical applications Sample preparation Cell separation Cell analysis Cell culture Molecular applications Reprogramming Embryonic and induced pluripotent stem cells Pioneering solutions for ES and iPS cells research Hematopoietic stem cells Making headway in your HSC research iPS cells Do you differentiate your ES or iPS cells? Characterization of reprogrammed cells Human • Extracellular PSC staining: TRA-1-60, TRA-1-81, SSEA-3, SSEA-4, CD326, CD324, etc. • Extracellular differentiated cell staining: SSEA-1 • Transcription factor staining: Oct4, Sox2, Nanog, etc. • MACSQuant® Flow Cytometers Mouse • Extracellular PSC staining: SSEA-1 • Extracellular differentiated cell staining: Anti-Feeder antibodies • Transcription factor staining: Oct4, Sox2, Nanog, etc. • MACSQuant Flow Cytometers Isolation of reprogrammed cells Magnetic isolation Human • Anti-TRA-1-60 MicroBead Kit • Pluripotent Stem Cell MicroBeads • Anti-SSEA-4 MicroBeads • Isolation of TRA-1-81+ cells via indirect magnetic labeling • hES Cloning and Recovery Supplement Mouse • Anti-SSEA-1 MicroBeads Flow sorting • Fluorochrome-conjugated antibodies (markers above) Colony Picking • StainAlive™ Antibodies Generation of iPS cells Human • mRNA Reprogramming Reagents • Pluriton™ Reprogramming Media • Small molecules Mouse • Reprogramming Ecotropic Retrovirus • Small Molecules Differentiation of ES or iPS cells Human or mouse • Cytokines • Small molecules Note: Different cytokines are used in accordance with the differentiation pathway. Purification of ES or iPS cells prior to differentiation Human • Anti-TRA-1-60 MicroBead Kit • Pluripotent Stem Cell MicroBeads Mouse • Pluripotent Stem Cell Isolation Kit Differentiation of ES or iPS cells via embryoid bodies • Embryoid Body Dissociation Kit • gentleMACS™ Dissociator Monitoring differentiation by flow cytometry or immunohistochemistry • Antibodies, for example, use CD34 antibodies to analyze cells during differentiation into hematopoietic or endothelial lineages. • MACSQuant Flow Cytometers Purification of cells during or after differentiation MicroBead reagents, for example, cell separation reagents for differentiated cells, e.g., • Neural Crest Stem Cell MicroBeads • CD34 MicroBeads • CD144 (VE-Cadherin) MicroBeads • CD309 (Flk-1) MicroBead Kit • Anti-PSA-NCAM MicroBead Kit • Anti-A2B5 MicroBead Kit Human samples Mouse samples Sample type Frozen • Dead Cell Removal Kit • Pre-Separation Filters PBMNCs or lysed whole blood • Red Blood Cell Lysis Solution Isolation of HSCs Magnetic isolation • Lineage Cell Depletion Kit + CD117 MicroBeads + Anti-Sca-1 MicroBead Kit (FITC) • CD105 Multisort Kit (PE) + Anti-Sca-1 MicroBead Kit (FITC) Flow sorting Fluorochrome-conjugated antibodies against, e.g., • Lineage markers • Anti-Sca-1 • CD117 (c-kit) • CD135 (Flt3R)Isolation of HSCs Magnetic isolation • CD34 MicroBead Kit • CD133 MicroBead Kit • Lineage Cell Depletion Kit • CD117 MicroBead Kit Flow sorting Fluorochrome-conjugated antibodies against, e.g., • CD34 • CD133 • CD45 • CD90 • CD38 Expansion of HSCs • Cytokines, e.g., TPO, SCF, Flt3 Expansion of HSCs • Cytokines, e.g., SCF, Flt3 Phenotyping of HSCs Human Fluorochrome-conjugated antibodies against, e.g., • CD34 • CD133 • CD45 • CD90 • CD38 • MACSQuant® Flow Cytometers Mouse Fluorochrome-conjugated antibodies against, e.g., • Lineage markers • Sca-1 • CD117 (c-kit) • CD135 (Flt3R) • MACSQuant Flow Cytometers Molecular analysis of HSCs • mRNA Isolation Kit and cDNA Synthesis Kit (for RT-PCR) • Protein Isolation Kits (for ChIP or normal IPs) • Microarray services • SuperAmp™ Technology • Bioinformatics • MultiMACS™ M96 Separator • Mitochondria isolation CFU assays for HSCs Human • HSC-CFU Media Differentiation of HSCs Human or mouse • Cytokines Note: Different cytokines are used in accordance with the differentiation pathway. Characterization of HSCs ES cells Transfection of ES or iPS cells Human • Stemfect™ hESC DNA Transfection Kit • Stemfect Enhanced hESC DNA Transfection Kit • Stemfect RNA Transfection Kit • MACSelect™ Products Mouse • Stemfect 2.0-mESC DNA Transfection Reagent • MACSelect™ Products Single-cell passaging of ES or iPS cells Human • hES Cloning and Recovery Supplement • Anti-TRA-1-60 MicroBead Kit • Pluripotent Stem Cell MicroBeads Monitoring of pluripotency by flow cytometry or immunohistochemistry Human • Extracellular PSC staining: TRA-1-60, TRA-1-81, SSEA-3, SSEA-4, CD326, CD324, etc. • Extracellular differentiated cell staining: SSEA-1 • Transcription factor staining: Oct4, Sox2, Nanog, etc. • MACSQuant® Flow Cytometers Mouse • Extracellular PSC staining: SSEA-1 • Extracellular differentiated cell staining: Anti-Feeder antibodies • Transcription factor staining: Oct4, Sox2, Nanog, etc. • MACSQuant® Flow Cytometers Translate research to clinical applications • GMP Cytokines • CliniMACS System • CliniMACS® Reagents, e.g., CD34, CD133 Removal of feeder cells Depleting feeder cells Human • Anti-Fibroblast MicroBeads Mouse • Feeder Removal MicroBeads Isolating pluripotent stem cells Human • Anti-TRA-1-60 MicroBead Kit • Pluripotent Stem Cell MicroBeads Mouse • Anti-SSEA-1 MicroBeads Culture of ES or iPS cells Feeder-based Human • C ytokines, e.g., FGF2 • CryoStem™ Freezing Medium • Small molecules Mouse • Cytokines, e.g., LIF • Small molecules Feeder-free Human • NutriStem™ Medium • Cytokines, e.g., FGF2 • Small molecules • CryoStem Freezing Medium Mouse • Cytokines, e.g., LIF • Small molecules

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Sample preparation Cell separation Cell analysis Cell culture Molecular applications Clinical applications Sample preparation Cell separation Cell analysis Cell culture Molecular applications Reprogramming Embryonic and induced pluripotent stem cells Pioneering solutions for ES and iPS cells research Hematopoietic stem cells Making headway in your HSC research iPS cells Do you differentiate your ES or iPS cells? Characterization of reprogrammed cells Human • Extracellular PSC staining: TRA-1-60, TRA-1-81, SSEA-3, SSEA-4, CD326, CD324, etc. • Extracellular differentiated cell staining: SSEA-1 • Transcription factor staining: Oct4, Sox2, Nanog, etc. • MACSQuant® Flow Cytometers Mouse • Extracellular PSC staining: SSEA-1 • Extracellular differentiated cell staining: Anti-Feeder antibodies • Transcription factor staining: Oct4, Sox2, Nanog, etc. • MACSQuant Flow Cytometers Isolation of reprogrammed cells Magnetic isolation Human • Anti-TRA-1-60 MicroBead Kit • Pluripotent Stem Cell MicroBeads • Anti-SSEA-4 MicroBeads • Isolation of TRA-1-81+ cells via indirect magnetic labeling • hES Cloning and Recovery Supplement Mouse • Anti-SSEA-1 MicroBeads Flow sorting • Fluorochrome-conjugated antibodies (markers above) Colony Picking • StainAlive™ Antibodies Generation of iPS cells Human • mRNA Reprogramming Reagents • Pluriton™ Reprogramming Media • Small molecules Mouse • Reprogramming Ecotropic Retrovirus • Small Molecules Differentiation of ES or iPS cells Human or mouse • Cytokines • Small molecules Note: Different cytokines are used in accordance with the differentiation pathway. Purification of ES or iPS cells prior to differentiation Human • Anti-TRA-1-60 MicroBead Kit • Pluripotent Stem Cell MicroBeads Mouse • Pluripotent Stem Cell Isolation Kit Differentiation of ES or iPS cells via embryoid bodies • Embryoid Body Dissociation Kit • gentleMACS™ Dissociator Monitoring differentiation by flow cytometry or immunohistochemistry • Antibodies, for example, use CD34 antibodies to analyze cells during differentiation into hematopoietic or endothelial lineages. • MACSQuant Flow Cytometers Purification of cells during or after differentiation MicroBead reagents, for example, cell separation reagents for differentiated cells, e.g., • Neural Crest Stem Cell MicroBeads • CD34 MicroBeads • CD144 (VE-Cadherin) MicroBeads • CD309 (Flk-1) MicroBead Kit • Anti-PSA-NCAM MicroBead Kit • Anti-A2B5 MicroBead Kit Human samples Mouse samples Sample type Frozen • Dead Cell Removal Kit • Pre-Separation Filters PBMNCs or lysed whole blood • Red Blood Cell Lysis Solution Isolation of HSCs Magnetic isolation • Lineage Cell Depletion Kit + CD117 MicroBeads + Anti-Sca-1 MicroBead Kit (FITC) • CD105 Multisort Kit (PE) + Anti-Sca-1 MicroBead Kit (FITC) Flow sorting Fluorochrome-conjugated antibodies against, e.g., • Lineage markers • Anti-Sca-1 • CD117 (c-kit) • CD135 (Flt3R)Isolation of HSCs Magnetic isolation • CD34 MicroBead Kit • CD133 MicroBead Kit • Lineage Cell Depletion Kit • CD117 MicroBead Kit Flow sorting Fluorochrome-conjugated antibodies against, e.g., • CD34 • CD133 • CD45 • CD90 • CD38 Expansion of HSCs • Cytokines, e.g., TPO, SCF, Flt3 Expansion of HSCs • Cytokines, e.g., SCF, Flt3 Phenotyping of HSCs Human Fluorochrome-conjugated antibodies against, e.g., • CD34 • CD133 • CD45 • CD90 • CD38 • MACSQuant® Flow Cytometers Mouse Fluorochrome-conjugated antibodies against, e.g., • Lineage markers • Sca-1 • CD117 (c-kit) • CD135 (Flt3R) • MACSQuant Flow Cytometers Molecular analysis of HSCs • mRNA Isolation Kit and cDNA Synthesis Kit (for RT-PCR) • Protein Isolation Kits (for ChIP or normal IPs) • Microarray services • SuperAmp™ Technology • Bioinformatics • MultiMACS™ M96 Separator • Mitochondria isolation CFU assays for HSCs Human • HSC-CFU Media Differentiation of HSCs Human or mouse • Cytokines Note: Different cytokines are used in accordance with the differentiation pathway. Characterization of HSCs ES cells Transfection of ES or iPS cells Human • Stemfect™ hESC DNA Transfection Kit • Stemfect Enhanced hESC DNA Transfection Kit • Stemfect RNA Transfection Kit • MACSelect™ Products Mouse • Stemfect 2.0-mESC DNA Transfection Reagent • MACSelect™ Products Single-cell passaging of ES or iPS cells Human • hES Cloning and Recovery Supplement • Anti-TRA-1-60 MicroBead Kit • Pluripotent Stem Cell MicroBeads Monitoring of pluripotency by flow cytometry or immunohistochemistry Human • Extracellular PSC staining: TRA-1-60, TRA-1-81, SSEA-3, SSEA-4, CD326, CD324, etc. • Extracellular differentiated cell staining: SSEA-1 • Transcription factor staining: Oct4, Sox2, Nanog, etc. • MACSQuant® Flow Cytometers Mouse • Extracellular PSC staining: SSEA-1 • Extracellular differentiated cell staining: Anti-Feeder antibodies • Transcription factor staining: Oct4, Sox2, Nanog, etc. • MACSQuant® Flow Cytometers Translate research to clinical applications • GMP Cytokines • CliniMACS System • CliniMACS® Reagents, e.g., CD34, CD133 Removal of feeder cells Depleting feeder cells Human • Anti-Fibroblast MicroBeads Mouse • Feeder Removal MicroBeads Isolating pluripotent stem cells Human • Anti-TRA-1-60 MicroBead Kit • Pluripotent Stem Cell MicroBeads Mouse • Anti-SSEA-1 MicroBeads Culture of ES or iPS cells Feeder-based Human • C ytokines, e.g., FGF2 • CryoStem™ Freezing Medium • Small molecules Mouse • Cytokines, e.g., LIF • Small molecules Feeder-free Human • NutriStem™ Medium • Cytokines, e.g., FGF2 • Small molecules • CryoStem Freezing Medium Mouse • Cytokines, e.g., LIF • Small molecules

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Stem Cell quick guide 2013

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Stem Cell quick guide 2013