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1 UK +44 (0) CH +41 (0) DE +49 (0) US

2 Featured Product Areas Stem Cell Fate Regulators and Synthetic Retinoid ec23 Recombinant Growth Factor Mimetics Stem Cell Differentiation Antibodies Adult Stem Cell Differentiation Media 3D Culture Specialized Reagents, Natural and Recombinant 3D Synthetic Scaffold Stem cells are unspecialized cells that are capable of dividing and renewing themselves for long periods of time and can differentiate into many specialized cell types. Differentiation of stem cells into the desired derivative represents a significant challenge. Ultimately, the goal is to produce robust and reliable stem cell differentiation processes that can be controlled within the laboratory. AMSBIO offers the most extensive range of differentiation factors, media, kits and tools for 3D culture. Differentiation Factors Stem Cell Fate Regulators MAPTrix Recombinant Factors Recombinant Factors 3D Culture Cultrex 3D BME, Collagen & Laminin MAPTrix HyGel Mimetics Alvetex Synthetic Scaffold Differentiation Kits and Media Marrow Stromal Stem Cell Differentiation Mesenchymal Stem Cell Differentiation Neural Stem Cell Differentiation Adipocyte Stem Cell Differentiation info@amsbio.com

3 Differentiation Factors The differentiated progeny of a stem cell is dictated by the environment and signals received. The in vitro differentiation tissue culture conditions can be tightly controlled by researchers. To promote differentiation of stem cells, signaling factors or inhibitors can be added to the tissue culture medium and/or the surface matrix on which the stem cells are in contact can be modified. To address the needs of today s stem cell researchers, AMSBIO supplies a wide range of differentiation factors. Stem cell fate regulator set II Stem Cell Fate Regulator Set II is a convenient set of eight small molecule modulators useful in stem cell research. The eight modulators are: BIX01294, CHIR99021, PD , PD , RG 108, SB , SU and Valproic Acid Sodium. Modulators are also sold separately. Synthetic retinoid ec23 ec23 is a small molecule that enables the robust and reproducible differentiation of stem cells and progenitor cells. It has a much higher stability within tissue culture medium and is a more potent differentiation factor than retinoic acid, resulting in the down regulation of markers associated with the pluripotent stem cell phenotype and increased expression of differentiation markers. A B C D Synthetic retinoid ec23 induced differentiation: ec23 is a potent inducer of neurogenesis in adult neuroprogenitor cells. Rat adult hippocampal progenitor cells exposed to 10µM ec23 differentate into neural cell types. Differentiation induced by exposure to ec23 was consistent and reproducible. Images show ec23 treated cells, phase contrast (A) and stained with β-tubulin (B). (C). Phase contrast micrograph of human TERA2.cl.SP12 EC stem cells. (D). Phase contrast micrograph of human TERA2.cl.SP12 EC cells treated for 28 days Note phase bright aggregations of neurons linked to each other by bundles of axons. Stem Cell Fate Regulator Set II K Set BIX mg CHIR mg PD mg PD mg RG mg SB mg SU µg Valproic Acid Sodium mg Synthetic retinoid ec23 Contact info@amsbio.com UK +44 (0) CH +41 (0) DE +49 (0) US

4 MAPTrix recombinant growth factor mimetics A growth factor is a naturally occurring substance capable of stimulating cellular growth, proliferation and cellular differentiation. MAPTrix Growth Factor mimetics offer a unique presentation of growth factor sequences to stem cells in culture. Small peptide mimics of natural growth factors have been incorporated into a recombinant adhesive protein backbone structure. This innovative product is biofunctional - growth factor mimetic sequences are presented to the stem cell while the adhesive backbone anchors the protein to the tissue culture surface. The presentation of growth factors from an anchored surface recapitulates the in vivo environment whereby growth factors are presented to cells by the extracellular matrix. MAPTrix Growth Factor Mimetics have comparable biological activity to soluble growth factor peptides. Animal-free MAPTrix recombinant mimetics are available for FGF, EGF, PDGF, VEGF and many more. This product is user-friendly and supplied in a ready-to-use formulation. Guaranteed high lot-to-lot consistency and reproducibility with QC & QA testing in accordance with current USP Guidelines. To find which MAPTrix products you need, check or contact info@amsbio.com MAPTrix Technologgy Mussel Adhesive Protein Economical Adhesion to a wide variety of surfaces Chemically Defined and Animal-Free No species specificity Adhere to USP guidelines Low endotoxin and no E.coli derived protein Tested negative for bacteria, fungi and mycoplasma Recombinant differentiation cytokines and factors An extensive range of recombinant proteins, growth factors and cytokines are available. Check or contact info@amsbio.com Stem cell research antibodies AMSBIO supplies a wide and varied range of antibodies supporting stem cell differentiation. Whether you require phenotype markers for undifferentiated stem cells or for the differentiated progeny, AMSBIO is your source for high quality stem cell research antibodies. Differentiation Medium and Kits AMSBIO supplies performance tested differentiation media and complete kits along with optimized protocols to ensure successful differentiation of marrow stromal, mesenchymal, adipose and neural stem cells every time. Marrow stromal stem cell adipocyte and osteoblast differentiation media Human Marrow Stromal Stem Cells provide a convenient in vitro model for investigating pathways of multipotent cell differentiation. Human Marrow Stromal Stem Cells can be effectively differentiated into Adipocytes and Osteoblasts with optimized protocols and differentiation media. Human Marrow Stromal Stem Cell Adipocyte Differentiation Medium SMH811D ml Human Marrow Stromal Stem Cell Osteoblast Differentiation Medium SMH417D ml info@amsbio.com

5 Mesenchymal stem cell differentiation to adipocytes and osteoblasts, kits and media Mesenchymal Stem Cells can be isolated from bone marrow by adherence to plastic and can differentiate into multiple lineage-specific cells that form bone, fat, cartilage, muscle and tendon. To facilitate reproducible and consistent evaluation of mesenchymal cell potential and research, a range of mesenchymal differentiation kits and mediums are available. Neural stem cell differentiation media Neural Stem Cells are multipotent and can be induced to differentiate into neurons, astrocytes and oligodendrocytes. Neural Stem Cell Differentiation Media are available supporting rat and mouse neural stem cell differentiation. Media and protocols to differentiate adult adipose derived stem cells into adipocytes, osteoblasts and chondrocytes Adult Stem Cells derived from adipose tissue are multipotent and can be induced differentiate into osteoblasts, chondrocytes or adipocytes upon exposure to specific differentiation media formulations. Differentiation media supporting differentiation of Human Adipose Derived Stem Cells are available for all three differentiation fates. Adult stem cells differentiated into adipocytes. Cells shown are two weeks old and have been stained with the neutral lipid dye Oil Red O. Adult stem cells differentiated into osteoblasts. Mineralized cells stained with Alizarin Red. Adult stem cells differentiated into chondrocytes. Cells stained with Alcian Blue. Rat Mesenchymal Stem Cell Adipocyte Differentiation Medium SR811D ml Rat Mesenchymal Stem Cell Osteoblast Differentiation Medium SR417D ml Mesenchymal Stem Cell Adipogenic Differentiation Kit K 1 Kit Mesenchymal Stem Cell Osteogenic Differentiation Kit K 1 Kit Rat Neural Stem Cell Differentiation Medium SMRD ml Mouse Neural Stem Cell Differentiation Medium SMMD ml Adult Adipose Derived Stem Cell Adipocyte Differentiation Medium DM-2 100ml Adult Adipose Derived Stem Cell Adipocyte Differentiation Medium DM ml Adult Adipose Derived Stem Cell Osteoblast Differentiation Medium OB-1 100ml Adult Adipose Derived Stem Cell Osteoblast Differentiation Medium OB ml Adult Adipose Derived Stem Cell Chondrocyte Differentiation Medium CM-1 100ml Adult Adipose Derived Stem Cell Chondrocyte Differentiation Medium CM-1-500ml 500ml UK +44 (0) CH +41 (0) DE +49 (0) US

6 3D Stem Cell Culture While it is possible to direct cell differentiation toward certain lineages in vitro, it is especially challenging to produce higher-order structures that are composed of more complex arrangements of cells which grow and differentiate together within a 3D space that more closely resembles the in vivo environment. AMSBIO offers the most comprehensive product solutions for the 3D culture of stem cells with natural and recombinant 3D matrices and synthetic scaffolds, quality tested and guaranteed pathogen free. Cultrex 3D Basement Membrane Extract, collagen I and laminin The Cultrex range includes 3D culture formats of Basement Membrane Extract (BME), Collagen I and Laminin that have been specifically qualified to promote 3D patterns of morphology, polarity and proliferation. Related products include 3D proliferation assays, cell harvesting kit and cell invasion assays. Stem Cell Qualified BME is also available for general application. Cultrex 3D Culture Matrix BME ml Cultrex 3D Culture Matrix BME ml Cultrex 3D Collagen I mg Cultrex 3D Culture Laminin I mg MAPTrix recombinant hydrogel ECM mimetics MAPTrix HyGel are semi-synthetic 3D extracellular matrices that are tailored to mimic specific ECM properties, without any animal derived components. These hydrogels are composed of two components: MAPTrix ECM mimetics, which are small peptide mimics of extracellular matrices fused with a recombinant adhesive protein and a multi-arm polyethylene glycol derivative linker. Easily engineer the elasticity or the pore size of the hydrogel by adjusting the concentration of MAPTrix ECM or MAPTrix Linker. Use of MAPTrix HyGel generates a well-controlled and reproducible cellular environment for your 3D cell culture and related application. The immobilized short peptide cell adhesion mimetic motifs are available for laminin, fibronectin, vitronectin and collagen. To find which MAPTrix products you need, check or contact info@amsbio.com MAPTrix Hygel- Laminin (4 motifs) MAPTrix ECM + MAPTrix Link = MAPTrix Hygel- Fibronectin (3 motifs) MAPTrix Hygel- Collagen (2 motifs) MAPTrix Hygel- Vitronectin (2 motifs) MAPTrix products are economical, convenient and highly reproducible with excellent lot-to-lot consistency with no animal derived components. MAPTrix mediated adipocyte differentiation: Mesenchymal stem cells from a male mouse adipose were encapsulated and cultured in MAPTrix Hygel TM Collagen type I for 7 days. The encapsulated MSCs were then transplanted to the abdominal cavity of a female mouse for one month before analysis. Y chromosome detection was used to identify male implanted cells. info@amsbio.com

7 Alvetex synthetic scaffold for 3D differentiation Traditionally, cultured cells have been grown on treated-polystyrene 2D surfaces as in standard cell culture plastic -ware. AMSBIO now offers the Alvetex synthetic scaffold that allows cells the equivalent growth and differentiation substrate in a 3D format. The Alvetex synthetic scaffold is a thin membrane made from inert polystyrene with a well defined and uniform micro-porous architecture providing consistent and reproducible results (see figure below). A B Alvetex is compatible with standard cell culture coating reagents : (A) The ECM protein forms a web of fibers spanning voids into which cells can differentiate in 3D. (A) Alvetex loaded with Collagen. (B) Alvetex coated with Basement Membrane Extract and seeded with human pluripotent stem cells and induced to differentiate. An acinus/gland-like structure with central lumen could be observed. Sample stained with Masson's Trichrome, picking out the blue of the collagen present in the Basement Membrane Extract. 3D cultures on Alvetex can be studied using a variety of standard molecular and cellular techniques, including: Tissue processing, fixation, embedding and sectioning Bright-field, fluorescence and confocal microscopy Electron Microscopy both SEM and TEM Histological staining and immunocytochemistry, In situ hybridization Extraction of nucleic acid and protein Laser capture 100µm Protein expression in 3D cell cultures can be localized using immunocytochemistry and fluorescence microscopy. Three corresponding images from the same region of Alvetex show:(a) cell morphology by phase microscopy; (B) expression of cell nucleus marker DAPI; (C) expression of cell proliferation marker Ki67. All Alvetex products are supplied as non-tissue culture treated. These 3D culture products can additionally be treated with any extracellular matrix in the same manner as traditional 2D cell culture plasticware. Alvetex is supplied as 12 well plates and as a cradle housing three 6-well inserts, developed to fit into a 90mm Petri dish, in order to accommodate larger media volumes that can support long term 3D cell culture. Alvetex products will be supplied as either non-tissue culture treated or tissue culture treated, and in a variety of other formats including standard size 6, 24, 48 and 96 well plates with clips and well inserts to fit 6, 12 and 24 well plates (coming soon). Contact info@amsbio.com with your specific requirements. Cultrex Stem Cell Qualified Basement Membrane Extract ml Alvetex 3D culture 12 well AMS.AVP002 (1 plate) 1 plate Trademarks are used here for the purposes of identification only. These trademarks are the property of their indicated owners as follows: ec23 and Alvetex Reinnervate Ltd.; MAPTrix and MAPTrix HyGel, Kollidis Biosciences Inc.; and Cultrex, Trevigen, Inc. UK +44 (0) CH +41 (0) DE +49 (0) US

8 AMS Biotechnology - info@amsbio.com Phone: UK +44 (0) CH +41 (0) DE +49 (0) US Fax: UK +44 (0) CH +41 (0) DE +49 (0) US AMSBIO - Cutting Edge Stem Cell Reagents and Kits! info@amsbio.com