biomedica newsletter - biomedica:...

BIOMEDICANEWSLETTER

MARCH 2010

Dear valued customer,

Research in developmental biology has led to the discovery of several cell types. Biomedica Medizinprodukte displays means to grow, analyze and manipulate cells and decipher molecular pathways, all equally significant in regenerative and deve-lopmental biology.

In this edition we present our range of products in the field of Stem Cell Culture and Cell Biology. Our suppliers in this promising research area are:

If you need any further information, do not hesitate to contact your customer advisor:

Sales Manager Life Science CZ / SK

RNDr. Jiří Brůna [email protected] +420 724370069

Sales Representative Life Science CZ

Martin Holub, PhD. [email protected] +420 725 819 186

Customers Sales Service CZ

Petra Darmovzalová

Pavla Theodossisová

[email protected] +420 283 933 616+420 283 931 485

fax: +420 283 932 507

Sales Representative Life Science SK

Mgr. Patrícia Gasperova [email protected] +421 911 750 208

Customers Sales Service SK

Maria Bizónova [email protected] +421 2 693 099 01

CONTENT

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R&D SYSTEMS Multicolor Flow Cytometry KitsMycoplasma Detection KitMesenchymal Stem Cell Functional ID KitMagCellectTM Mouse/Human NK Cell Isolation KitHuman Pluripotent Stem Cell Ab Array

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PROMOCELL Preadipocyte & Adipocyte CultureStem- and Blood- Cells, PericytesCD133+ Progenitor CellsFetal Calf Serum, MSC/ESC-QualifiedAngiogenesis ModelsMycoplasma Detection & Elimination

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PROMOKINECell Transfection

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ABGENTStem Cell ResearchAntibody Sampler Kits For Stem Cells

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PANOMICSProcarta Profiling KitsQuantiGene Plex 2.0 Reagent System

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SPECIAL OFFERS

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tBIOMEDICA NEWSLETTER MARCH 2010 LIFE SCIENCE

IMPRESSUM: BIOMEDICA CS s.r.o, Na Safrance 1651/9, Praha 101 00, Czech Republic, www.biomedica.cz

BIOMEDICA SK s.r.o., Drobného 27, Bratislava 841 01, Slovakia, www.biomedica.sk

The multicolor flow cytometry kits from R&D Systems contain antibodies conjugated to up to four different fluorochromes: phycoerythrin (PE), allophycocyanin (APC), Peridinin Chlo-rophyll Protein Complex (PerCP), and Carboxyfluorescein (CFS). The application of antibody-specific fluorochromes allows simultaneous assessment of up to four different target molecules. Washed cells are fluorescently stained, with the

intensity of staining directly proportional to the expression of the target molecule. All multicolor flow cytometry kits in-clude isotype controls for each antibody, providing specific experimental comparisons.Multicolor flow cytometry kits are available for hematopoi-etic stem cells, Th17 cells, and multipotent mesenchymal stromal cells.

R&D SYSTEMS

Multicolor Flow Cytometry Kits

Fig. 1: Th17 4-Color Flow Cytometry Kit. Human Peripheral Blood Mononuclear Cells (PBMCs) were stained with the human Th17 4-Color Flow Cytometry Kit (RD-FMC007). Dot plots show IL-17+ cells in relation to CD3+, IL-23 R+, or IL-22+ populations from activated PBMCs. Quadrants were set based on isotype controls.

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Mycoplasma Detection Kit of R&D Systems represents a ma-jor breakthrough for high-throughput, routine screening of cell cultures for mycoplasma contamination. Mycoplasma con-tamination is one of the most common and serious problems in culturing eukaryotic cells because it alters the phenotypic characteristics of the cells and can negatively impact results.

Since mycoplasma is typically not visible and does not respond to antibiotics, an alternative sensitive and reliable detection method is required. This assay is capable of detecting the eight mycoplasma species known to cause 95% of eukaryotic cell culture contamination. For more details you can visit the R&D website or consult your Biomedica sales contact.

Mycoplasma Detection Kit

R&D Systems is a recognized source for high quality stem cell reagents, kits, markers, and more. With a profoundly or-ganized website and detailed catalogues one easily finds the specific products for used techniques as well as developed reagents for the in vitro culture, manipulation, and charac-terization of stem cells. The broad and permanently growing

product palette provides opportunities to study multiple as-pects of embryology and offers unprecedented potential for the disease treatment. The newcomers are released almost daily. In order to keep close track on the new products cus-tomers can register free of charge to receive a newsletter at: http://www.rndsystems.com/request_catalog.aspx

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BIOMEDICA NEWSLETTER MARCH 2010 LIFE SCIENCE

Human Pluripotent Stem Cell Antibody Array

Fig. 4: The Human Pluripotent Stem Cell Array detects multiple protein mark-ers in various cell extracts.

The Human Pluripotent Stem Cell Antibody Array is a rapid, sensitive, and economical tool to simultaneously detect the relative expression of 15 pluripotent stem cell markers in a single sample. No specialized equipment is necessary.

Mesenchymal Stem Cell Functional Identification Kit

MagCellect™ Mouse and Human Natural Killer Cell Isolation Kits

Fig. 3: Enrichment of NK cells from Peripheral Blood Mononuclear Cells (PB-MCs) using the MagCellect Human NK Cell Isolation Kit (RD-MAGH109).

This R&D Systems kit is designed to identify bone marrow-derived stem cells (BMSCs) and mesenchymal stem cells (MSCs) based on their ability to differentiate into multiple mesenchymal lineages. The kit contains specially formulated media supplements that can be used to effectively differen-tiate BMSCs/MSCs into adipocytes, osteocytes, and chon-drocytes. A panel of antibodies including goat anti-mouse FABP4, mouse anti-human Osteocalcin, and goat anti-hu-man Aggrecan are also provided to assess the mature phe-notypes of each lineage. Available specific for human, rat and mouse mesenchymal stem cells.

The MagCellect NK Cell Isolation Kits are designed to isolate NK cells via a process of negative selection that removes unwanted cells from a mononuclear cell suspension. The kits contain an NK cell biotinylated antibody cocktail, streptavidin ferrofluid, and buffer with sufficient reagents to process 1 x 109 cells. The typical purity of the recovered NK cells ranges between 70-80% for the mouse kit and 80-90% for the hu-man kit. Following selection, the enriched cell preparation can be used for a variety of applications including tissue cul-ture, immune status monitoring, and flow cytometry.

Fig. 2: Differentiation and Identification of Rat Mesenchymal Stem Cells (MSCs) using the Rat Mesenchymal Stem Cell Functional Identification Kit (RD-SC020)

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PROMOCELL

Preadipocyte & Adipocyte Culture

Since PromoCell was established in 1990 in Heidelberg, this company has always endeavoured to support the trend towards carrying out in vitro cell culture tests using normal human primary cells. They are a responsible partner in all ar-

Growth of the adipose tissue results from both the enlarge-ment of mature adipocytes and the formation of new adipo-cytes from adipocyte precursor cells (preadipocytes). The dif-ferentiation process of adipocyte precursor cells is controlled by a variety of hormones. Primary cultures of preadipocytes provide a useful approach for the investigation of physiologi-cal mechanisms controlling cell proliferation, differentiation and maintenance of function in adipose tissue. Substantial

progress in the culture of preadipocytes and adipocyte dif-ferentiation process has recently been achieved with the development of a serum-free differentiation medium. This is important due to the inhibitory effects of serum on the differ-entiation of preadipocytes. PromoCell´s adipocyte cell culture system provides preadi-pocytes combined with optimised growth and differentiation media.

Fig. 5: Left: Undifferentiated Human White Preadipocytes (HWP) using phase contrast. Middle: HWP after in vitro differentiation into adi-pocytes using the PromoCell Preadipocyte Differentiation Medium (phase contrast). Right: HWP after in vitro differentiation into adipocytes (Lipid droplets stained with Oil Red O).

eas of cell culture research and associated specialist fields.They are now the leading European manufacturer in the field of primary human cell culture.

Stem- and Blood- Cells

PromoCell offers a collection of adult stem cells as well as differentiated blood cells from normal human bone marrow, umbilical cord tissue, adipose tissue, peripheral blood, and cord blood. All cells are obtained from approved donor pro-grams at associated medical centers. High quality media that simulate the in vivo environment as well as optimized dif-

ferentiation media systems are also available for these cell types.

The PromoCell Stem- and Blood- Cells are isolated, as all other PromoCell Cells, according to strict ethical guidelines.

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Pericytes are multipotent mesenchymal-like cells found in association with small blood vessel walls. They are impor-tant for angiogenesis, the structural integrity of the microvas-culature, and blood flow regulation. However, they can also develop into malignant tumors called hemangiopericytomas.So far, these cells have demonstrated the ability to differenti-ate into adipocytes, osteoblasts, fibroblasts, smooth muscle cells, and phagocytes (macrophages).

Fig. 6: Proliferating human pericytes (hPC-PL) from the placenta (phase contrast).

CD133 (AC133, Prominin-1) is a glycoprotein with sub-cellu-lar localization in plasma membrane protrusions. In humans, this protein was originally identified as an antigenic marker expressed on hematopoietic stem and progenitor cells. In addition, CD133 is also expressed by neural and muscle stem cells. CD133+ cells are isolated in PromoCell´s facili-

ties from fresh cord blood. These cells are highly proliferative primitive progenitors. They can give rise to mature terminally differentiated hematopoietic cells by means of directed dif-ferentiation. PromoCell also provides suitable expansion and differentiation media.

Fetal Calf Serum, MSC-Qualified

The PromoCell MSC-Qualified Fetal Calf Serum is thorough-ly pre-tested for its efficiency in isolation and expansion of Mesenchymal Stem Cells (MSC) derived from bone marrow, umbilical cord matrix and adipose tissue. The rigid quality

controls performed in the PromoCell stem cell lab stand for maximum viability and straightforward expansion of MSCs while maintaining their full differentiation capacity.

Fig. 7: Expansion and differentiation of Mesenchymal Stem Cells (MSC) using media with the PromoCell MSC- Qualified Fetal Calf Serum. Left: Undifferentiated hMSC-BM cultured for 4 passages (phase contrast). Right: The same cells after 14 days of adipogenic induction (Oil Red O staining of lipid droplets with Hema-toxylin counterstain).

CD133+ Progenitor Cells

Pericytes NEW

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The PromoCell ESC-Qualified Fetal Calf Serum is thorough-ly pre-tested for culture of Embryonic Stem Cells (ESC). The rigid quality controls performed in the PromoCell stem cell lab stand for maximum expansion of unaltered ESCs while maintaining unrestricted pluripotency.

Fetal Calf Serum, ESC-Qualified

Fig. 8: Growth morphology of murine embryonic stem cells (mESC) in stan-dard serum vs. PromoCell ESC- Qualified Fetal Calf Serum. Left: mESC in standard FCS. Note the large, flat, differentiated cells. Right: mESC after 5 passages in PromoCell ESC- Qualified Fetal Calf Serum. The cells are small and have a compact colony morphology typical for mESC.

Angiogenesis Models

Angiogenesis, the growth of new blood vessels from pre-existing ones, is a normal process in growth and develop-ment, as well as in wound healing. However, it is also vitally involved in a number of pathological processes like tumor growth and metastasis, rheumatoid arthritis, and macular degeneration. It occurs in four well-characterized stages, which are controlled by pro-angiogenic and anti-angiogenic factors:

Proteolytic activity: Angiogenic growth factors activate • receptors on endothelial cells in pre-existing blood ves-sels. The activated endothelial cells begin to release proteases that degrade the basement membrane.Migration: Endothelial cells from the parent vessel walls • migrate through the gaps in the basement membrane into the surrounding tissue.Proliferation: The migrated endothelial cells proliferate • and form solid sprouts. These sprouts extend toward the source of the angiogenic stimulus.Lumen formation: The cells in the solid sprouts form a • lumen, which results in a fully developed vessel.

PromoCell offers four diverse angiogenesis models to simu-late these angiogenesis stages or the whole process in vitro, thus making pro- or anti-angiogenic testing fast and effi-cient:

3D Angiogenesis Assay• Planar Migration Assay• Transmigration Kit• Invasion Kit•

Fig.9: stimulated / unstimulated Endothelial Cell Spheroids in the 3D- Angio-genesis- Assay

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Frequency of Testing:

New cell and virus cultures• Each month for continuous cell lines; each week in • cases of laboratory contamination

Mycoplasma Detection and Elimination

Before each liquid nitrogen storage• Upon modification of the cell characteristics• In case of problems with result reproducibility•

PCR Mycoplasma Test Kit I/C – conventional PCR technologyPCR Mycoplasma Test Kit I/RT – real time PCR technology

Features:

Highly sensitive (detects 10 – 15 mycoplasmas per • sample volume)Fast, reproducible, and reliable results• Applicable for all cell/tissue culture-relevant myco-•

plasma speciesPositive and internal controls included• Convenient protocol and easy sample preparation•

Mycoplasma elimination in cell/tissue culture and virus stocks.

Mycoplasma detection in cell culture, cell/ virus stocks, and mediums.

Features:

Highly effective against M. orale, M. hyorhinis, M. homi-• nis, Acholeplasma laidlawii, M. arginini, M. fermentans, and other mycoplasma species usually encountered as contaminants in cell cultures combined non-antibiotic and antibiotic treatment • ease of use – relatively fast procedure• temperature stable, ready-to-use solution • no additional dosing during the treatment necessary • kills mycoplasmas efficiently - but is safe for cells• Fig. 10: Mycoplasma contaminated Cell Culture from: www.unc.edu/depts/

tcf/mycoplasma.htm

Mycoplasma-EX

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PROMOKINEa brand of PromoCell

The ability to introduce foreign nucleic acids into eukaryotic cells has become a powerful tool in studying and controlling gene expression. PromoKine’s PromoFectin Transfection Reagents are ideal for highly effective and reliable transfection of a variety of cell types, including many hard-to-transfect cell lines, neuronal cells, sensitive primary cells and insect cells. PromoFectin can be used for optimal delivery of plasmid DNA, oligonucle-otides, siRNA and proteins/peptides into adherent and non-adherent cells. Transfecting cells using PromoFectin requires no medium change and shows very low cytotoxicity. Magnet Assisted Transfection (MATra) is an easy-to-handle and extremely fast technology to optimize transfection of

Cell Transfection

cells in culture. Exploiting a strong magnetic force, the full nucleic acid dose bound to magnetic nanoparticles is rapidly drawn towards the target cells resulting in a very efficient transfection. All types of nucleic acids (e.g. plasmid DNA, siRNA, oligonucleotides) can be used with the MATra ap-proach. MATra shows low cytotoxicity, is functional with and without serum and is very time-saving and cost-effective. MATra can also be adapted to high-throughput transfection assays using robotic stations and adapted protocols.

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Fig. 11: Primary human hepatocytes transfected with the reporter vector pPK-CMV-R4 using PromoFectin- Hepatocyte.

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ABGENT

Visit the website of Abgent: http://www.abgent.com/ to get de-tailed information about their broad range of products.Abgent develops primary antibodies also providing custom peptide synthesis and antibody development. The products

mainly focus on research topics such as Autophagy, Apopto-sis, Stem Cells, Phosphorylation and Cell Function & Gene Regulation research. In addition synthesis of difficult peptides including long peptides, is a core competency of Abgent.

Stem cell research

Fig.12: Formalin-fixed and paraffin-embedded human lung carcinoma tissue reacted with SOX2 monoclonal antibody.

Abgent offers more than 1000 antibodies specific for stem cell research, including popular targets such as AC133, ALDH, Oct4, c-KIT, and NANOG, as well as emerging tar-gets from the latest cutting edge research. The EpishotTM Antigen Prediction Screen enables Abgent to design optimal peptides for stem cell targets.

Sampler kits offer an affordable way to purchase multiple validated antibodies from Abgent’s comprehensive antibody product lines. Assemble a collection of antibodies (minimum of 5 antibodies per order) to your pathway of interest, or eco-nomically screen for the antibody that is perfect for your ap-plication.

Biomedica offers now products from Abgent!

Antibody Sampler Kits for Stem Cells

Fig.13: LEFT:Formalin-fixed and paraffin-embedded human hepatocarcinoma tissue reacted with AC133 (CD133) antibody (C-term), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. RIGHT:Immunofluorescence analysis of anti-AC133 (CD133) Antibody (C-term) in HeLa cells. 0.025 mg/ml primary antibody was followed by Alexa-Fluor-546-conjugated donkey anti-rabbit lgG (H+L). Alexa-Fluor-546 emits orange fluorescence. Blue counterstaining is DAPI.

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Procarta Cytokine assays can simultaneously quantitate up to 30 human cytokines from diverse matrices in 3 hours with a sensitivity of 10 pg/mL/cytokine. Human, Mouse, and Rat Cytokine Assay Kits are available in 1- and 10- plate sizes.

PANOMICS

Plexable Human Cytokine, Chemokine and Growth Factors (50 Plex)EGF IFN alpha IL-5 IL-15 MCP-3 PDGF BB TNF betaENA-78 IFN beta IL-6 IL-17A MCSF Rantes VEGFEotaxin IFN gamma IL-7 IL-17F MIG ResistinFGF basic IL-1 alpha IL-8 IL-20 MIP-1 alpha SAAGCSF IFN-beta IL-10 IP10 MIP-1 beta sCD40 ligandGMCSF IL-1 ra IL-12 (p40) ITAC MIP-3 alpha TGF betaGRO alpha IL-2 IL-12 (p70) Leptin NGF TGF alphaHGF IL-4 IL-13 MCP-1 PAI-1 TNF alpha

Additional Plexable AnalytesThese 33 analytes can be plexed with the above 50 targets.Adiponectin Granzyme B IL-16 MIF SDF-1 TPOBLC ICAM IL-21 MPO sE-Selectin TRAILCTACK IFN omega IL-22 PIGF sFas VCAMEotaxin 2 IL-2 ra IL-23 SAP sFAS LigandEotaxin 3 IL-3 IL-31 SCF sRANKLFractlakine IL-9 LIF SCGF-b Survivin/BIRC5

Panomics, now part of Affymetrix, offers a broad variaty of Luminex bead based arrays.

Procarta Cytokine Profiling Kits

Human MMP Panel MMPs can be plexed with our Human Cytokine/Chemokine assaysMMP-1 MMP-2 MMP-3 MMP-7MMP-8 MMP-9 MMP-12 MMP-13

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Procarta Transcription Factor Profiling Kits

Quantitate the DNA binding activity of up to 44 different transcription factors (TFs) in a single well using nuclear extracts or whole cell lysates.

Procarta SH2 Domain Plex Profiling Kits

Profile phosphotyrosine protein interactions against 30 SH2 binding in a single well using treated and untreated cell lysates.

QuantiGene Plex 2.0 assays combine branched DNA (bDNA) signal amplification technology and xMAP® (multi-analyte profiling) beads to enable simultaneous quantification of multiple RNA targets directly from cultured cell or whole blood lysates; fresh, frozen or formalin-fixed, paraffin-embedded (FFPE) tissue homogenates; or purified RNA preparations.

QuantiGene Plex 2.0 Reagent System

Fig.14: Panomics Procarta Transcription Factor Assay was used to simultaneously measure the activity of 40 TFs in a single assay well. HeLa cells were serum starved overnight and subsequently stimulated PMA or a vehicle control for 4 hours. As expected, PMA induced AP1 and NF-E2 and the activity of NF-1 and STAT4 were unchanged. Sample inductions from the Procarta TF assay were confirmed by EMSA Gel Shift assays. Lane 1 = Free Probe; Lane 2 = HeLa Nuclear Extract; and Lane 3 = PMA Induced HeLa Nuclear Extract.

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3D-Angiogenesis Assay UNIQUEThe PromoCell ready-to-use 3D-Angiogenesis Assay allows fast and easy screening for pro-angiogenic and anti-angiogenic effects of test substances in a convenient 24-well format. The assay simulates the entire angiogenesis process in vitro in three dimen-sions.

Until April 30, 2010, you get a 30% discount on your order of a 3D-Angiogenesis Assay!Please use code PC1/10 when ordering.

Promotions

Don’t Break My Heart - Get Cardiac Cells from PromoCell instead!PromoCell offers high quality cells from the cardiac system including: Human Cardiac Myocytes, Human Cardiac Fibroblasts, and Human Cardiac Microvascular Endothelial Cells.

Until April 30, 2010, with your order of these cells you will get 1 bottle of the recomend-ed PromoCell Medium free of charge.Please use code PC1/10 when ordering.

For further promotion details please contact your Biomedica sales representative!

BIOMEDICA CS s.r.o.Na Safrance 1651/9

Praha 101 00Czech Republic

BIOMEDICA SK s.r.o.Drobného 27

Bratislava 841 01Slovakia

Cell Transfection:Receive a 15% discount on all PromoKine Cell Transfection products from 1st of March until 31st of May 2010. Please use code SP-PK-0310 when ordering.

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2010

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