Stem Cells by Miltenyi Biotec

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miltenyibiotec.com/stemcells Embryonic and induced pluripotent stem cells Pioneering solutions for ES and iPS cells research

Differentiation Reprogramming

Purification of ES and iPS cells prior to differentiation Fibroblasts isolation

Human Mouse • Whole Skin Dissociation Kit • TRA-1-60 MicroBeads • Pluripotent • gentleMACS™ Dissociators • Feeder Removal Isolation Kit MicroBeads • Feeder Removal • Anti-Fibroblast MicroBeads Generation of reprogrammed cells MicroBeads • StemMACS mRNAs for reprogramming and differentiation Induction of differentiation • StemMACS mRNA Transfection Reagent • StemMACS Small Molecules • MACS • StemMACS™ Small Molecules, such as CHIR99021, Purmorphamine, and more Isolation of reprogrammed cells • StemMACS mRNA, including Brn2, Mash1 Human Mouse Note: Different reagents are used in accordance • Anti-TRA-1-60 • Anti-SSEA-1 MicroBeads with the differentiation pathway. MicroBeads • Flow sorting using • Anti-SSEA-4 MicroBeads the MACSQuant Tyto Transfection and transduction of ES and iPS cells • StemMACS Y27632 • StemMACS Thiazovivin • MACSelect™ Products • Flow sorting using • StemMACS mRNA Transfection Kit the MACSQuant Tyto • StemMACS Nuclear eGFP mRNA • MACSfectin™

Monitoring differentiation by flow cytometry Maintenance or Culture of ES and iPS cells • MACS Portfolio Example application: Use CD34 Human Mouse to analyze cells during differentiation into • StemMACS iPS-Brew XF • MACS Cytokines, such hematopoietic or endothelial lineages. • StemMACS Passaging as LIF • MACSQuant® Flow Cytometers Solution XF • StemMACS™ Small • StemMACS Small Molecules, such as Purification of cells during or after differentiation Molecules, such as CHIR99021, PD0325901 Y27632, Thiazovivin, Removal of remaining ES or iPS cells CHIR99021, PD0325901 • UEA-1 MicroBeads • MACS Cytokines, such as FGF-2 Ectoderm • Anti-TRA-1-60 MicroBeads • Neural Crest Stem Cell MicroBeads • Anti-A2B5 MicroBeads Kit • Anti-PSA-NCAM MicroBead Kit Phenotyping of ES and iPS cells • CD133 MicroBead Kit Endoderm Human Mouse • CD184 (CXCR4) MicroBead Kit MACSQuant Flow MACSQuant Flow Cytometers and Cytometers and Mesoderm fluorochrome-conjugated fluorochrome- • CD34 MicroBead Kit antibodies for detection of conjugated antibodies for • CD144 MicroBeads • Pluripotency: TRA-1-60, detection of: • CD309 (Flk-1) MicoBead Kit TRA-1-81, SSEA-4, • Pluripotency marker, CD326, CD324 such as SSEA-1, CD326 • Differentiated cells: • Feeder cells: SSEA-1 Anti-Feeder • Feeder cells: Anti-Feeder, Anti-Fibroblast

Sample preparation Cell separation Flow cytometry Cell culture Molecular applications Clinical applications Hematopoietic stem cells Making headway in your HSC research

Sample preparation

• Dead Cell Removal Kit • Smart Strainers • Pre-Separation Filters • Red Blood Cell Lysis Solution

Human samples Mouse samples

Isolation of HSCs Isolation of HSCs

• CD34 MicroBead Kit UltraPure • Lineage Cell Depletion Kit • CD133 MicroBead Kit – Hematopoietic Tissue • Anti-Sca-1 MicroBead Kit (FITC) • CD117 MicroBead Kit • CD117 MicroBeads • Lineage Cell Depletion Kit • CD105 MultiSort Kit (PE) • Flow sorting using the MACSQuant Tyto and your favorite • Flow sorting using the MACSQuant Tyto and your favorite marker panel marker panel

Culture of HSCs Culture of HSCs

Expansion Expansion • StemMACS HSC Expansion Media XF • MACS Cytokines, including SCF, Flt3-, Il-3, Il-6 • StemMACS HSC Expansion Cocktail Differentiation • MACS Cytokines, including TPO, SCF, Flt3-Ligand, Il-3, Il-6 • MACS Cytokines, such as Il-7, GM-CSF Differentiation • StemMACS HSC-CFU Media • MACS Cytokines, such as Il-7, GM-CSF

Phenotyping and flow cytometry Phenotyping and flow cytometry

MACSQuant Flow Cytometers in combination with MACSQuant Flow Cytometers in combination with fluorochrome-conjugated MACS Antibodies including: fluorochrome-conjugated MACS Antibodies, such as • CD34 • Lineage markers • CD133 • Sca-1 • CD45 CHECK OUT OUR • CD117 (c-) • CD38 EXCLUSIVE VIO®DYES! • CD135 (Flt3) • CD90

Molecular anaylsis of HSCs Translate research to clinical applications • MACS GMP Cytokines • µMACS™ mRNA Isolation Kit • SuperAmp™ Technology – cDNA • CliniMACS® and CliniMACS Prodigy® • µMACS Isolation Kits from just a few cells • CliniMACS Reagents, including CD133 (epitope tags, protein A/G) • Magnetic mitochondria isolation • MultiMACS™ M96 Separator • Enhanced viral transduction with MACSductin

StemMACS HSC Expansion Media XF • Xeno-free expansion of hematopoietic stem and progenitor cells • Consistently high expansion rates • High CD34+ cell content after expansion Mesenchymal stem cells Driving your MSC research

Sample preparation

• Dead Cell Removal Kit • Smart Strainers • Pre-Separation Filters • gentleMACS Dissociators

Human samples Mouse samples

Phenotyping or flow sorting of MSCs prior to culture Phenotyping or flow sorting of MSCs prior to culture

Fluorochrome-conjugated antibodies against, for example: Fluorochrome-conjugated antibodies against, for example: • CD271 • CD146 • Lineage markers • CD45 • MSCA-1 • CD105 • Sca-1 • CD105

Isolation of MSCs prior to culture Isolation of MSCs prior to culture

MicroBeads for positive isolation MicroBeads for removal of non-MSCs • CD271 MicroBead Kit • CD146 MicroBead Kit • Lineage Cell Depletion Kit • Anti -Ter119 MicroBeads • Anti-MSCA-1 (W8B2) • CD105 MicroBeads • CD45 MicroBeads • CD11b MicroBeads MicroBead Kit • Anti-Fibroblast MicroBeads

Culture of MSCs Culture of MSCs

Expansion Differentiation • MACS Cytokines, including TGF-β, EGF • StemMACS MSC • StemMACS AdipoDiff Expansion Media Kit XF • StemMACS ChondroDiff • StemMACS MSC • StemMACS OsteoDiff Phenotyping of expanded MSCs Expansion Media • CytoMix – MSC • Fluorochrome-conjugated antibodies against, for example, • MACS Cytokines, such as lineage markers, CD105, CD117, Anti-Sca-1, CD45, CD116, FGF-2, PDGF-BB, TGF-β and more

Phenotyping of expanded MSCs

• MSC Phenotyping Kit • Fluorochrome-conjugated antibodies against, for example, CD73, CD90, CD105, CD45, CD34, CD14, CD20, and more

Molecular anaylsis of MSCs Translate research to clinical applications • MACS GMP Cytokines • µMACS mRNA Isolation Kit • SuperAmp™ Technology – cDNA • CliniMACS and CliniMACS Prodigy • µMACS Protein Isolation Kits from just a few cells • CliniMACS CD271 Reagent (epitope tags, protein A/G) • Magnetic mitochondria isolation • MultiMACS™ M96 Separator • Enhanced viral transduction with MACSductin™

StemMACS MSC Expansion Media Kit XF • Xeno-free expansion of human mesenchymal stem cells • No attachment substrate required • Excellent expansion rates Embryonic and induced pluripotent stem cells Highlighted products

StemMACS iPS-Brew XF

• Xeno-free culture system for human ES and iPS cells • Easy transition to feeder-free conditions • Formulation compliant with future clinical translation

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StemMACS™ Small Molecules

• Highly pure for consistent biological activity • Pre-tested on stem cell cultures • Detailed instructions for preparation of stock solutions

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StemMACS™ mRNA Technology

• Modified mRNAs for modulation of cell fate by mRNA transfection • Optimized sequences, validated expression • Validated protocol for mRNA reprogramming

Download the mRNA reprogramming manual

Human FGF-2 IS

• Improved sequence for increased stability in cell culture • Standardized, high biological activity • Produced in E. coli, free of animal components

See the data Fore more information visit www.miltenyibiotec.com/stemcells

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Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use. CliniMACS, gentleMACS, MACS, MACSductin, MACSfectin, MACSQuant, MACSelect, MultiMACS, Prodigy, StemMACS, SuperAmp, and μMACS are either registered trademarks or trademarks of Miltenyi Biotec GmbH. CryoStem, Pluriton, StainAlive, and Stemfect are all trademarks of Stemgent, Inc.

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