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Tissue regener research br

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  • 1. Table of contentsMACS® Products for tissue regeneration research 3MACS Technology—the complete solution 4MACS Cell Separation Strategies 5Human stem cells—sample preparation, cell separation and analysis 6Embryonic stem cells and induced pluripotent stem cells—separation and analysis 7Mesenchymal stromal cells—separation and analysis 8Endothelial progenitor cells—separation and analysis 9Tissue regeneration—examples 10References 12MACS Products for sample preparation, cell separation, and cell analysis 13MACS Products for cell culture 14Molecular biology products and services 15Cover image: SPL/Agentur Focus
  • 2. MACS® Technology MACS® Products for tissue regeneration research MACS® Products provide the basis for reliable methods to investigate the full potential of stem MACS Products provide the tools that open up new cells in tissue regeneration research. perspectives for tissue regeneration research: Stem and progenitor cells have the potential to revolutionize • Reliable isolation of defined stem cell populations tissue regeneration and engineering in the future. The with excellent purity for reproducible results understanding of stem cell biology is of paramount importance • Stem cells and progenitors thereof can be isolated for the development of stem cell–based therapies. The within an hour rather than several days, multipotent—and even pluripotent—differentiation capacity saving valuable time for you to focus on your research. of certain stem and progenitor cell populations makes them a potential key to successful regeneration of many tissue types. • Isolated cells can immediately be used for cell culture Over the coming decade, research into this field may take center and animal experiments; cells remain viable and fully stage as the new frontier in the treatment of many disabling functional. diseases and injuries. • Miltenyi Biotec offers integrated solutions for a wide Reproducible data, standardized protocols, and defined variety of research needs—from sample preparation cell sources are key to unravel the full potential of stem and to molecular analysis. progenitor cells. However, most stem cell sources consist of heterogenous cell populations with varying differentiation and regeneration potential. The availability of defined cell populations with specific differentiation capacities will therefore be crucial for the targeted regeneration of tissues. In turn, this will facilitate a controlled and optimized growth and differentiation of stem cells into their target tissue types, especially during ex vivo tissue development. The separation of defined pre-committed stem and progenitor cell populations derived from stem cell populations with pluripotent or multipotent differentiation potential would be beneficial for future tissue regeneration applications. Miltenyi Biotec offers numerous innovative products for the isolation of stem and progenitor cells as well as for sample preparation, cell analysis, cell culture, and molecular analysis. 3
  • 3. MACS® Technology MACS® Technology —the complete solution Since its introduction in 1989, MACS® Technology has become the gold standard for cell separation. Nowadays, Miltenyi Biotec stands for more than cell separation, offering more than 1000 innovative research products for biomedical research and life sciences. The MACS Research Product portfolio includes instruments and reagents for sample preparation, cell separation, cell analysis, cell culture, and molecular biology. Miltenyi Biotec also provides tools for clinical-scale cell separation based on MACS Technology. Miltenyi Biotec has a strong commitment to continual product development with regards to current and future basic and clinical research. MACS Sample Preparation The quality of an experiment strictly depends on the quality of the sample preparation. Miltenyi Biotec offers innovative instruments and reagents for fast and gentle preparation of samples from solid tissues as well as cultured cells. MACS Cell Separation A large panel of MACS MicroBeads and MicroBead Kits is available for the isolation of virtually any cell type. The cells can be separated manually or in an automated fashion. The new autoMACS™ Pro Separator has been designed for automated walk-away cell sorting of multiple samples. MACS Cell Analysis Miltenyi Biotec provides a large panel of monoclonal antibodies and kits for fluorescence microscopy and flow cytometry. The innovative MACSQuant™ Analyzer is an extremely compact, easy-to-use, multicolor benchtop cell analyzer. The instrument is fully automated and enables absolute cell count. MACS Cell Culture The product portfolio for cell culture includes media as well as recombinant cytokines and growth factors. MACSmolecular Miltenyi Biotec provides products for analytical protein isolation and detection, mRNA purification and amplification, cDNA synthesis and labeling, microRNA analysis, as well as microarray technologies and instrumentation. The portfolio includes genomics services, such as gene expression and microRNA analysis microarrays, array-CGH, and bioinformatics.4
  • 4. MACS® Cell Separation Strategies MACS® Technology—the gold standardBenefits of MACS® Technology at a glance: in cell separation• Fast—cell separation takes less than one hour MACS Technology is based on MACS MicroBeads, MACS• Gentle—separated cells remain viable and functional Columns, and MACS Separators—strong permanent magnets.• Flexible—both labeled and unlabeled fractions can be MicroBeads are superparamagnetic particles coupled to specific obtained with excellent purity and high recovery monoclonal antibodies.• Easy separation of large cell numbers—up to 109 Target cells can be magnetically isolated by positive selection labeled cells per column using specific cell surface antigens or by depletion of unwanted cells in order to obtain untouched cells. Furthermore, these two separation strategies can easily be combined to provide greaterMACS Technology flexibility for the sequential sorting of complex subpopulations Magnetic labeling of cells. Cells of interest are labeled with MACS® MicroBeads in a short MACS MicroBeads incubation step. • Highly specific monoclonal antibody conjugates • Small (50 nm), virus-sized nanoparticles • Non-toxic, biodegradable Magnetic separation Labeled and unlabeled cells are • Colloidal, for ease of handling and short incubation times separated on a MACS Column placed in the magnetic field of a MACS Columns and MACS Separators MACS Separator. • Optimal recovery and high purity with MACS Columns The flow-through can be collected • Gentle to cells as the non-magnetic, unlabeled cell fraction. • Automated cell separation with autoMACS™ Pro Separator Elution of the labeled cell fraction The separation column is removed from the magnetic field and the retained cells are flushed out. Both the labeled and unlabeled fractions can be recovered and used for downstream applications. MidiMACS™ Separator autoMACS™ Pro Separator MACS MicroBeads for indirect magnetic labeling For maximum flexibility, indirect magnetic labeling with MACS MicroBeads allows the use of any primary antibody. Monoclonal or polyclonal primary antibodies can be either unconjugated, biotinylated, or fluorochrome-conjugated. MACS MicroBeads are nano-sized particles and are barely detectable by scanning electron microscopy. The micrograph shows a lymphocyte isolated by positive selection (left). Transmission electron micrograph of an isolated lymphocyte with MicroBeads (arrow) on the cell surface (right). (Courtesy of Prof. Groscurth, Zürich, CH.) 5
  • 5. Sample preparation,cell separation, cell analysis Human stem cells—sample preparation, cell separation and analysis Human stem cells in tissue regeneration research Stem and progenitor cells have the potential to revolutionize tissue regeneration and engineering in the future. To support research into the biology of stem cells and to explore their extraordinary capacity for future tissue regeneration applications, it is crucial to develop reliable methods for the specific isolation of distinct stem and progenitor cell populations with defined differentiation potential. Likewise, the isolation of their differentiated progenies is of immense interest for research into future clinical applications. For stem cell research, Miltenyi Biotec has developed numerous innovative tools that allow straightforward sample preparation, cell separation, as well as subsequent cell analysis. Sample preparation For a broad range of tissues that are of interest for current tissue regeneration research, the ideal cell sources have yet to be determined. Therefore, the isolation of stem cells from embryonic, fetal, and adult sources is of great importance. Preparing suitable suspensions of viable single cells from tissues is a critical step for the successful isolation of tissue-specific stem cells. Miltenyi Biotec offers the new gentleMACS™ Dissociator for the gentle and efficient dissociation of tissues. In addition, the Neural Tissue Dissociation Kits, Pre-Separation Filters, and the Dead Cell Removal Kit allow an optimal preparation of single-cell suspensions for subsequent separation. gentleMACS™ Dissociator The gentleMACS™ Dissociator is a benchtop instrument for the automated dissociation of tissues. Two types of unique gentleMACS Tubes are available for the instrument and enable the time-saving and easy dissociation of tissues into single-cell suspensions or homogenization of tissues to lysates in a closed system. The range of protocols available for the gentle and efficient dissociation of tissues is continuously expanding. Protocols are optimized to yield single-cell suspensions with high viability rate. The gentleMACS Dissociator can further be used for the homogenization of tissues or cells to extract biomolecules for molecular biology experiments, such as the isolation of total RNA or mRNA. gentleMACS Dissociator—features at a glance • Time-saving automated tissue dissociation or homogenization The gentleMACS Dissociator—the gentle way of automated tissue dissociation • Standardized procedure • Reliable and reproducible results • High level of user safety • Sterile sample handling Cell separation and analysis For research into future tissue regeneration applications, it is crucial to develop reliable methods for the specific isolation of distinct stem and progenitor cell populations with defined differentiation potential as well as their differentiated progenies. Miltenyi Biotec offers numerous tools for the isolation of stem and progenitor cells according to specific cell surface markers. For maximum flexibility, indirect magnetic labeling with MACS MicroBeads allows the use of any primary antibody. Monoclonal or polyclonal primary antibodies of choice can be either unconjugated, biotinylated, or fluorochrome-conjugated. Furthermore, a large variety of products is available for the analysis of stem and progenitor cells.6
  • 6. Cell separation and analysis Embryonic stem cells and induced pluripotent stem cells— separation and analysis Embryonic stem cells and induced pluripotent stem cells Pluripotent embryonic stem cells (ESCs), derived from the inner cell mass of the blastocyst, and induced pluripotent stem (iPS) cells are known for their capacity to differentiate into virtually all cell types of the body and to self-renew while maintaining a stable karyotype. Therefore, these cells are of special interest for basic tissue regeneration research to understand developmental processes, and might provide the basis for clinical applications Cell separation in the future. For these applications it is crucial to use homogenous cell populations. MACS® Cell analysis Technology enables the isolation of numerous particular cell types and subsets at high microRNA expression profiling purity—ready for in vitro and animal experiments. Gene expression analysis Pluripotent cells express the surface markers SSEA-3, SSEA-4, Tra-1-60, Tra-1-81, but not CD15 (SSEA-1).1–3 The positive selection, for example, according to the expression of SSEA-4 using MACS MicroBeads for indirect magnetic labeling is a proven strategy for the isolation of pluripotent cells.4,5 These markers can also be used for the depletion of pluripotent cells from differentiated cultures, thereby reducing the risk of teratoma formation in transplantation experiments. For more information on Miltenyi Biotec products for ESC and iPS cell research, please refer to the After their in vitro differentiation, ESC- or iPS-derived cells exhibit a broad cellular corresponding brochure that is available for download at www.miltenyibiotec.com. heterogeneity with respect to developmental stages and lineage specification. This leads to highly heterogenous cell populations. Cell separation according to the expression of certain surface markers can enrich target cells or deplete unwanted populations: Hematopoietic stem cells (HSCs) can be isolated from differentiated ESCs by positive selection according to the expression of, for example, CD34 or CD133.6–12 Endothelial progenitor cells (EPCs) can be separated from ESC cultures by using, for example, the cell surface markers CD347,8,13 and CD31. Early neural cell populations were enriched with FORSE-1 antibodies and MicroBeads for indirect magnetic labeling. Various markers that indicate the developmental maturity after neuronal differentiation have been described: CD133+, CD271+, or CD146+ cells showed characteristics of neural stem or precursor cells, A2B5+ cells showed characteristics of glial-restricted precursor cells, whereas CD56+, PSA- NCAM+, or CD24+ cells showed characteristics of differentiated neurons.14 Cardiomyocytes might be separable from heterogenous ESC cultures according to CD56+ expression.15 7
  • 7. Cell separation and analysis Mesenchymal stromal cells—separation and analysis Mesenchymal stromal cells Mesenchymal stromal cells (MSCs) can be obtained from a variety of tissue sources including bone marrow aspirate16,17, umbilical cord blood17, and even lipoaspirate18,19. MSCs have shown the potential to differentiate into a variety of nonhematopoietic tissue types, including bone or cartilage. Numerous cell surface antigens have been used for the isolation of MSCs from various sources. Sample preparation Cell separation Bone marrow Cell analysis CD271 is a well-known marker for the isolation of MSCs from bone marrow. 20–22 Cell culture Marrow stromal antigen-1 (MSCA-1), which is recognized by clone W8B2, was shown to be Expression profiling restricted to MSCs in the CD271bright population in bone marrow.22 These CD271brightCD45dim MSCs show a higher clonogenic capacity compared to the CD271+CD45+ fraction.22 Therefore, MSCA-1 is a suitable marker for the identification of MSCs with a high proliferative potential. Additional markers for the isolation of MSCs from bone marrow include CD117 23, CD105 24,25, For more information on Miltenyi Biotec Stro-1 26 , CD146 27, and CD13328. products for MSC research, please refer to the corresponding brochure available for download at www.miltenyibiotec.com. Multipotent adult progenitor cells (MAPCs) can be enriched from bone marrow by depletion of CD235a (glycophorin A) and CD45. 33 Cord blood BM-MNCs before separation CD133 is a suitable marker for the identification of MSCs from cord blood.17 Lipoaspirate Various strategies can be pursued for the isolation of MSCs from lipoaspirate: positive CD45-FITC selection of CD271+ cells 29 or CD146+ cells, positive selection of CD34+ cells30, as well as depletion of CD45+ and CD31+ cells. 18,31 Dental pulp STRO-1 and CD146 are appropriate markers for the separation of MSCs from dental pulp. 27 Anti-MSCA-1 (W8B2)-APC MSCA-1+ cells Amniocentesis cultures MSC-like fetal-derived stem cells from amniocentesis cultures can be isolated according to the expression of CD117 and share many markers of adult and embryonic stem cells. Fetal- derived stem cells retain a multipotent ability to differentiate into cell types that represent CD45-FITC all embryonic germ layers. 32 Anti-MSCA-1 (W8B2)-APC MSCA-1 (W8B2)+ cells were isolated from human bone marrow mononuclear cells (BM-MNCs) using the MSC Research Tool Box – MSCA-1 (W8B2). Cells were stained with Anti-MSCA-1 (W8B2)-APC and CD45-FITC.8
  • 8. Endothelial progenitor cells—separation and analysis Endothelial progenitor cells Regeneration of vascular tissue is an important topic in therapeutic research, especially for the potential treatment of peripheral vascular disease and the revascularization of ischemic tissues, for example, in the heart. Endothelial progenitor cells (EPCs) have been suggested to play an important role in postnatal neoangiogenesis and neovascularization. Therefore, EPCs have come into focus for the potential treatment of ischemic or injured tissue and for the coating of scaffolds to increase biocompatibility of biomaterials. EPCs are defined by the expression of the markers CD34 and CD309 (VEGFR-2/KDR). Analysis of CD133 expression allows the distinction between early and matured EPCs in human. 34,35 EPCs were enriched according to their expression of CD34 or CD133 from different hematopoietic sources34–38,43. CD133+ cells have been used in studies that show a significantly improved vascularSPECT (single-photon emission computed tomographic) scan from the posteriorwall area of a human heart. Courtesy of Prof. Gustav Steinhoff, Rostock, Germany. network restoration in an ischemic hind limb rat model38 and an ischemic heart mouse model39. Furthermore, CD133+ cells were used in combination with biodegradable scaffolds for the three- dimensional tissue engineering of microvessels.40 Safety and feasibility and, moreover, efficacy was shown in several clinical trials investigating CD133+ cell therapy combined with coronary artery bypass grafting41,42 and cell therapy alone43. 9
  • 9. Tissue regeneration research Tissue regeneration research—examples Tissue-resident stem cells have been found in almost any tissue. A A better understanding of their biology and the development of methods for their isolation and expansion might therefore be useful for tissue regeneration. In addition, stem cells from the hematopoietic system have been described to contribute to the regeneration of tissues. Following, a few examples for research on the regeneration of certain tissues are presented. Liver tissue In contrast to other organs, the liver is known for its capacity to regenerate in situ. This is due to the population of stem cells within the liver. These stem cells have been successfully isolated using MACS® Technology by targeting CD326 (EpCAM)44,45, CD13345, or CD117.46 The transplantation of bone marrow–derived B CD133 + cells has been shown to benefit the expansion of liver tissue in situ prior to partial hepatectomy.47 Muscle tissue The identification and isolation of muscle stem and progenitor cells by the markers CD13348 and CD5649,50, respectively, is of importance for research on smooth, skeletal, and even cardiac muscle tissue regeneration. After transplantation into mouse models, human CD133 + stem cells from peripheral blood and circulating endothelial progenitor cells (cEPCs) from umbilical cord blood were capable of regenerating in situ dystrophic CD133+ cells, isolated from mobilized peripheral blood, gave rise to adherent cells muscle tissue51 and skeletal muscle52, respectively. Cardiac after 3–5 weeks of cultivation. muscle regeneration is a prime goal in cardiovascular disease These cells were able to differentiate into hepatocyte-like cells. (A) The cells are stained for hepatocyte nuclear factor-3 (FITC), albumin (Cy3), and research; functional cardiomyocytes can be generated in vitro nuclei (DAPI). (B) The cells are stained for cytokeratin 19 (Cy3) and nuclei (DAPI) (200×). from ESCs15, CD133 + fetal liver cells53, and from CD34 + cEPCs from (Courtesy of Selim Kuçi, Tübingen, Germany.) peripheral blood54. CD133+ cells isolated from mobilized PBMCs were cultivated for 3–5 weeks. Adherent cells were able to differentiate into skeletal muscle–like cells. The cells are stained for desmin (FITC), actin (Cy3), and nuclei (DAPI)(200×). (Courtesy of Selim Kuçi, Tübingen, Germany.)10
  • 10. Tissue regeneration research—examples Neural tissue A The regeneration of neural tissue has far-reaching consequences for the potential treatment of debilitating neurodegenerative diseases or injuries, including stroke and spinal cord damage. Human neural stem cells that were isolated from fetal brain according to CD133 expression have been shown to differentiate in vitro and in vivo into cells with neural phenotypes and even restore the function of damaged spinal tissue in mice. 55 Furthermore, CD133 + cells isolated from mobilized peripheral blood56 or skin57 can be differentiated into neural lineages. Neuronal-committed precursors from mammalian brain can be magnetically isolated for research purposes via the depletion of A2B5 + cells followed by the positive selection of PSA-NCAM+ cells.58 B CCD133+ cells isolated from mobilized PBMCs were cultivated for 3–5 weeks.Adherent cells were able to differentiate into neural-like cells.(A) Astrocyte-like cells stained for GFAP (Cy3), EPO (FITC), and nuclei (DAPI).(B) Oligodendrocyte-like cells stained with GFAP (FITC), MBP (Cy3), and DAPI(nuclei).(C) Neuronal-like cells stained for betatubulin III (Cy3) and nuclei (DAPI)(200×).(Courtesy of Selim Kuçi, Tübingen, Germany.)For more information on Miltenyi Biotecproducts for neuroscience research, please referto the corresponding brochure that is availablefor download at www.miltenyibiotec.com. 11
  • 11. ReferencesMACS® Technology References 1. Kaufman, D. S. et al. (2001) Proc. Natl. Acad. Sci. USA 98: 10716–10721. 2. Chang, K. et al. (2006) Blood 108: 1515–1523. 3. Carpenter, M. K. et al. (2003) Cloning Stem Cells 5: 79–88. 4. Cheng, L. et al. (2003) Stem Cells 21: 131–42. 5. Schulz, T. C. et al. (2004) Stem Cells 22: 1218–1238. 6. Bandi, S. and Akkina, R. (2008) AIDS Res. Ther. 5: 1. 7. Wang, Z. Z. et al. (2007) Nat. Biotechnol. 25: 317–318. 8. Chen, T. et al. (2007) Stem Cells 25: 392–401. 9. Narayan, D. A. et al. (2006) Blood 107: 2180–2183. 10. Galic, Z. et al. (2006) Proc. Natl. Acad. Sci. USA 103: 11742–11747. 11. Kaufman, D. S. et al. (2001) Proc. Natl. Acad. Sci. USA 98: 10716–10717. 12. Vodyanik, M. A. et al. (2006) Blood 108: 2095–2105. 13. Ferreira, L. S. et al. (2007) Circ. Res. 101: 286–294. 14. Pruszak, J. et al. (2007) Stem Cells 25: 2257–2268. 15. Xu, S. et al. (2006) Stem Cells and Dev. 15: 631–639. 16. Jones, E. A et al. (2002) Arthritis Rheum. 46: 3349–3360. 17. Tondreau, T. et al. (2005) Stem Cells 23: 1105–1112. 18. Boquest, A. C. et al. (2005) Mol. Biol. Cell 16: 1131–1141. 19. Meyerrose, T. E. et al. (2007) Stem Cells 25: 220–227. 20. Jones, E. A. et al. (2006) Cytometry B Clin. Cytom. 70: 391–399. 21. Quirici, N. et al. (2002) Exp. Hematol. 30: 783–791. 22. Bühring, H. J. et al. (2007) Ann. NY Acad. Sci. 1106: 262–271. 23. Huss, R. and Moosmann, S. (2002) Br. J. Hematol. 118: 305–312. 24. Aslan, H. et al. (2006) Stem Cells 24: 1728–1737. 25. Majumdar, M. K. et al. (2003) J. Biomed. Sci. 10: 228–24. 26. Gronthos, S. et al. (1994) Blood 84: 4164–4173. 27. Shi, S. and Gronthos, S. (2003) J. Bone Miner. Res. 18: 696–704. 28. Pozzobon, M. et al. (2008) Stem Cells and Dev. (in press). 29. Godthardt, K. (2007) MSC 2007 Adult Mesenchymal Stem Cells in Regenerative Medicine, Cleveland, Poster Nr. 301, Session III. 30. Astori, G. et al. (2007) J. Transl. Med. 5: 55. 31. Noer, A. et al. (2006) Mol. Biol. Cell 17: 3543–3556. 32. De Coppi, P. et al. (2007) Nat. Biotechnol. 25: 100–106. 33. Reyes, M. et al. (2002) J. Clin. Invest. 109 : 337–346. 34. Peichev, M. et al. (2000) Blood 95: 952–958. 35. Rafii, S. and Lyden, D. (2003) Nat. Med. 9: 702–712. 36. Gehling, U. et al. (2000) Blood 95: 3106–3112. 37. Taguchi, A. et al. (2004) J. Clin. Invest. 330–338. 38. Suuronen, E. et al. (2006) Circulation. 114 (suppl. 1): 138–144. 39. Ma, N. et al. (2006) Cardiovasc. Res. 71: 158–169. 40. Wu, X. et al. (2004) Am. J. Physiol. Heart. Circ. Physiol. 287: H480–H487. 41. Stamm, C. et al. (2004) Thorac. Cardiovasc. Surg. 52: 152–158. 42. Stamm, C. et al. (2007) J. Thorac. Cardiovasc. Surg. 133: 717–725. 43. Klein, H. M. et al. (2007) Heart Surg. Forum 10: E66–69. 44. Schmelzer, E. et al.(2007) J. Exp. Med. 204: 1973–1987. 45. Schmelzer, E. et al. (2006) Stem Cells 24: 1852–1858. 46. Laurson, J. et al. (2005) Int. J. Exp. Pathol. 86: 1–18. 47. Schulte am Esch, J. et al. (2005) Stem Cells 23: 463–470. 48. Alessandri, G. et al. (2004) Lancet 364: 1872–1883. 49. Sinanan, A. et al. (2004) Biotechnol. Appl. Biochem. 40: 25–34. 50. De Luna, N. et al. (2006) J. Biol. Chem. 281: 17092–17098. 51. Torrente, Y. et al. (2004) J. Clin. Invest. 114: 182–195. 52. Pesce, M. et al. (2003) Circ. Res. 93: 51–62. 53. Schmelkov, S. V. et al. (2005) Circulation 111: 1175–1183. 54. Badorff, C. et al. (2003) Circulation 107: 1024–1032. 55. Cummings, B. J. et al. (2005) Proc. Natl. Acad. Sci. USA 102: 14069–14074. 56. Kuçi, S. et al. (2003) MACS&more 7/1: 6–8. 57. Belicchi, M. et al. (2004) J. Neurosci. Res. 77: 475–486. 58. Seidenfaden, R. et al. (2006) Mol. Cell Neurosci. 32: 187–198.12
  • 12. MACS® Products MACS® Products for sample preparation, cell separation, and cell analysis Product Components or capacity Order no. Product Order no. gentleMACS™ gentleMACS™ Dissociator 130-093-235 Anti-FITC MicroBeads 130-048-701 Starting Kit C Tubes, 25 pieces Anti-PE MicroBeads 130-048-801 M Tubes , 25 pieces Power cord Anti-APC MicroBeads 130-090-855 User manual gentleMACS Protocols Anti-Cy5/Anti-Alexa Fluor 647 MicroBeads 130-091-395 C Tubes C Tubes, 25 pieces, single- 130-093-237 Anti-Cy7 MicroBeads 130-091-652 packed Anti-Biotin MicroBeads 130-090-485 M Tubes M Tubes, 25 pieces, single- 130-093-236 Streptavidin MicroBeads 130-048-101 packed 130-093-458 M Tubes, 50 pieces per bag Rat Anti-Mouse IgG1 MicroBeads 130-047-101 Neural Tissue 50 tests with up to 400 mg of 130-092-628 Rat Anti-Mouse IgG2a+b MicroBeads 130-047-201 Dissociation Kit (P) neural tissue Rat Anti-Mouse IgM MicroBeads 130-047-301 Neural Tissue 50 tests with up to 400 mg of 130-093-231 Goat Anti-Mouse IgG MicroBeads 130-048-401 Dissociation Kit (T) neural tissue Mouse Anti-Rat Kappa MicroBeads 130-047-401 Pre-Separation 50 filters 130-041-407 Filters Goat Anti-Rat IgG MicroBeads 130-048-501 Dead Cell For 10⁹ total cells 130-090-101 Goat Anti-Rabbit IgG MicroBeads 130-048-602 Removal Kit MACS Products for indirect magnetic labeling MACS Products for sample preparation Product Order no. Product Order no. Anti-A2B5 MicroBeads, human, mouse, rat 130-093-388 Anti-A2B5-PE, -APC, pure, human, 130-093-581, 130-093-582, mouse, rat 130-092-394 MSC Research Tool Box – MSCA-1 (W8B2), 130-093-572 human Anti-MSCA-1 (W8B2)-FITC, -PE, -APC, 130-093-585, 130-093-587, -Biotin, pure, human 130-093-589, 130-093-593, Anti-MSCA-1 (W8B2) MicroBead Kit, human 130-093-583 130-093-595 Anti-PSA-NCAM MicroBeads, 130-092-966 Anti-PSA-NCAM-PE, -APC, human, 130-093-274, 130-093-273 human, mouse, rat mouse, rat CD31 MicroBead Kit, human 130-091-935 CD31-FITC, -PE, -APC, human 130-092-64, 130-092-653, CD34 MicroBead Kit, human 130-046-702 130-092-652 CD34 MultiSort Kit, human 130-056-701 CD34-FITC, -PE, -APC, human 130-081-001, 130-081-002, 130-090-954 CD45 MicroBeads, human 130-045-801 CD56-PE, APC, pure, human 130-090-755, 130-090-843, CD56 MicroBeads, human 130-050-401 130-090-955 CD105 MicroBeads, human 130-051-201 CD117 (A3C6E2)-PE, -APC, human 130-091-734, 130-091-733 CD117 MicroBead Kit, human 130-091-332 CD117 (AC126)-PE, human 130-091-735 CD133 MicroBead Kit, human 130-050-801 CD133/1 (AC133)-PE, -APC, -Biotin, 130-080-801, 130-090-826, pure, human 130-090-664, 130-090-422 CD146 MicroBead Kit, human 130-093-596 CD133/2 (293C3)-PE, -APC, -Biotin, 130-090-853, 130-090-854, CD235a (Glycophorin A) MicroBeads, human 130-050-501 pure, human 130-090-852, 130-090-851 MSC Research Tool Box – CD271 (PE), human 130-092-867 CD133/1 (W6B3C1) pure, human 130-092-395 MSC Research Tool Box – CD271 (APC), human 130-092-291 CD146-FITC, -PE, -APC, -Biotin, pure, 130-092-851, 130-092-853, CD271 MicroBead Kit (PE), human 130-092-819 human 130-092-849, 130-092-852, 130-092-850 Lineage Cell Depletion Kit, human 130-092-211 CD271 (LNGFR), -PE, -APC, -Biotin, 130-091-885, 130-091-884, CD271 MicroBead Kit (APC), human 130-092-283 human 130-091-883 CD326 (EpCAM) MicroBeads, human 130-061-101 CD309 (VEGFR/KDR)-PE, -APC, -Biotin, Coming soon human MACS Products for the isolation of stem and progenitor cells CD326 (EpCAM)-FITC, -PE, -APC, 130-080-301, 130-091-253, human 130-091-254 EPC Enrichment and Enumeration Kit, 130-093-477 human MACS Products for the analysis of stem and progenitor cells 13
  • 13. ProductsMACS® Technology MACS® Products for cell culture NH media for the enumeration, expansion, and MACS® NH Media enable the differentiation of mesenchymal stromal cells (MSCs) into functional nonhematopoietic (NH) cell types. differentiation of MSCs MSCs are of great importance to researchers working towards the development of novel tissue regeneration therapies. However, in order to properly evaluate the potential of MSCs completely, it is crucial to establish standardized and reproducible procedures for stem cell isolation and cultivation. MACS® NH Stem Cell Media are optimized for the most convenient enumeration and expansion of nonhematopoietic (NH) stem cells from a variety of sources, including bone marrow aspirate, lipoaspirate, or potentially any NH stem cell source, MSC enumeration e.g., bone marrow, NH CFU-F Medium tissue or organ within the human body. Miltenyi Biotec also lipoaspirate offers media to evaluate the differentiation potential of NH stem cells during in vitro cultivation. Media are available for the reliable and reproducible differentiation of NH stem cells to adipocyte, chondrocyte, or osteoblast lineages. These media can also be used to define the full differentation capacity of an NH stem cell population: MSCs must be able to form all three cell lineages. MSC expansion NH Expansion Medium MACS NH Media Order no. NH CFU-F Medium 24×5 mL 130-091-676 NH Expansion Medium 500 mL 130-091-680 NH AdipoDiff Medium 100 mL 130-091-677 NH ChondroDiff Medium 100 mL 130-091-679 NH OsteoDiff Medium 100 mL 130-091-678 CytoMix – MSC, human Adipocytes Chondrocytes Osteoblasts The CytoMix – MSC, human (130-093-552) is a composition of NH AdipoDiff Medium NH ChondroDiff Medium NH OsteoDiff Medium cytokines for the most efficient and reproducible expansion of human MSCs. In combination with the NH Expansion Medium, CytoMix – MSC optimally supports the proliferation of human MSCs, especially after separation, e.g., according to CD271 or MSCA-1 (W8B2) expression using MACS® Technology. MACS Basic Culture Media Order no. Basic media and cytokines for stem cell culture Miltenyi Biotec offers cell culture media for a broad spectrum DMEM 500 mL 130-091-437 of applications. In addition, high-quality recombinant cytokines DMEM with stable glutamine 500 mL 130-091-438 and growth factors are available, that are well-suited for various RPMI 1640 500 mL 130-091-440 applications such as cell culture, differentiation studies, and RPMI 1640 with stable glutamine 500 mL 130-091-439 biological assays. Selected products are available in a premium- grade format with excellent purity and high, well-defined activity MACS Cytokines and Growth Factors as well as in research-grade quality. For a complete list of available cytokines, please visit Human: BDNF; BMP-2; EGF; EG-VEGF; FGF-2; FGF-4; Flt3-Ligand; G-CSF; GM-CSF; HGF; IL-3; IL-6; IL-11; PDGF-AA; PDGF-AB; PDGF-BB; SCF; http://www.miltenyibiotec.com/cytokines. SDF-1α; TGF-α; TGF-β1; TGF-β3; TNF-α; VEGF (121 aa); VEGF (165 aa) Mouse: EGF; Flt3-Ligand, G-CSF; GM-CSF; IL-3 (135 aa); IL-6; SCF; TNF-α; VEGF (164 aa); VEGF (165 aa); Rat: VEGF-C; VEGF-C (C152S)14
  • 14. Molecular biology products and services Stem cell differentiation tracking by gene expression profiling MACSmolecular provides a highly innovative range of products and services with a strong focus on gene expression profiling. Particularly when isolating stem cells, sensitive downstream analyses are required. One-step mRNA isolation and in-column cDNA synthesis Premium mRNA is isolated within 15 minutes directly froma-Hyb™ Hybridization Station cells or tissues. The µMACS™ One-step cDNA Kit combines efficient magnetic isolation of mRNA with revolutionary in- column cDNA synthesis. Purified cDNA can be generated mRNA isolation/cDNA synthesis from just a few to as many as 10⁷ cells. µMACS mRNA Isolation Kit Small Scale 130-075-201 Large Scale 130-075-101 For Total RNA 130-075-102 PIQOR™ Stem Cell Microarray The PIQOR™ Stem Cell Microarray comprises 942 relevant marker µMACS mRNA Isolation Starting Kit 130-075-202 genes for human stem cells and their differentiation. µMACS One-step cDNA Kit 130-091-902 It is available as a convenient microarray kit* or within the scope µMACS One-step cDNA Starting Kit 130-091-989 of the microarray service**. Gene expression experiments allow for the quality control of different stem cell types, comparison PIQOR Microarray Kit * between different stages of differentiation, as well as the PIQOR Stem Cell Microarray Kit, antisense optimization of differentiation protocols. 4 Microarrays 130-092-033 SuperAmp™ Service *** 8 Microarrays 130-092-034 When the number of stem cells for analysis is low, Miltenyi Biotec PIQOR Microarray Service ** offers the ideal solution for gene expression profiling needs. The SuperAmp™ Service (available as an extension of Service Stem Cell Microarray Plus Amplification 160-000-765 the PIQOR™ Microarray Service) can reliably amplify mRNA SuperAmp Amplification *** million-fold from as little as one cell. The service is ideal for stem cells sorted with MACS® Technology, flow cytometry, SuperAmp Service (per sample) 160-000-936 or even from laser capture microdissected tissue. miRXplore Microarray Kit miRXplore™ Kits and Services 4 Microarrays 130-093-254 Explore microRNA expression in human and mouse stem 8 Microarrays 130-093-272 cells with the new miRXplore™ Microarray Kits and Services. Designed in collaboration with experts at the Rockefeller miRXplore Microarray Services University¹, the microarray covers more than 2700 human, miRXplore Microarray Service 160-001-143 mouse, rat, and viral microRNA sequences and possess rigorous miRXplore Universal Reference Service 160-001-161 internal control system. Sequences differing by just one oligonucleotide can be reproducibly detected and re-ratios miRXplore Additional Total RNA Extraction 160-001-162 calculated with the use of the proprietary miRXplore* PIQOR™ Microarray Kits are not available in the US and Canada. Universal Reference.** Microarray Service includes all experimental steps from RNA isolation to primary data analysis. Final data are returned including an extensive written report. Further Bioinformatics Services, such as pathway or cluster analysis, are Reference also available.*** In combination with the Microarray Services only. The SuperAmp Service 1. Landgraf, P. et al. (2007) Cell 129: 1401–1414. is not available for microRNA amplification 15
  • 15. Miltenyi BiotecMiltenyi Biotec was founded in 1989; nowadays, more than 1100employees develop, produce, and sell innovative products forcell research and clinical applications. Due to an intense focus onstem cell research at Miltenyi Biotec, the portfolio includes thelargest range of products available for the separation of stemcells. Moreover, Miltenyi Biotec offers integrated solutions for awide variety of research needs—from sample preparation tomolecular analysis.Miltenyi Biotec GmbH Miltenyi Biotec Inc. Miltenyi Biotec B.V. (Benelux) Miltenyi Biotec SAS (France) Miltenyi Biotec Asia Pacific 130-094-269Friedrich-Ebert-Straße 68 12740 Earhart Avenue macs@miltenyibiotec.nl Phone +33 1 56 98 16 16 Pte. Ltd. (Singapore)51429 Bergisch Gladbach Auburn, CA 95602, USA Customer service Netherlands macs@miltenyibiotec.fr Phone +65 6238 8183Germany Phone 800 FOR MACS, Phone 0800 4020120 macs@miltenyibiotec.com.sg Miltenyi Biotec S.r.l. (Italy)Phone +49 2204 8306-0 +1 530 888 8871 Customer service Belgium Phone +39 051 646 0411 Miltenyi Biotec S.L. (Spain)Fax +49 2204 85197 Fax +1 530 888 8925 Phone 0800 94016 macs@miltenyibiotec.it Phone +34 91 512 12 90macs@miltenyibiotec.de macs@miltenyibiotec.com Customer service Luxembourg macs@miltenyibiotec.es Phone 800 24971 Miltenyi Biotec K.K. (Japan) Miltenyi Biotec Phone +81 3 5646 8910 Miltenyi Biotec Ltd. (UK) Australia Pty. Ltd. Miltenyi Biotec Trading macs@miltenyibiotec.jp Phone +44 1483 799 800 Phone +61 02 8877 7400 (Shanghai) Co., Ltd. (P.R. China) macs@miltenyibiotec.co.uk macs@miltenyibiotec.com.au Phone +86 21 6235 1005www.miltenyibiotec.com macs@miltenyibiotec.com.cnUnless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use.MACS is a registered trademark of Miltenyi Biotec GmbH. a-Hyb, μMACS, MidiMACS, autoMACS, gentleMACS, MACSQuant, miRXplore, PIQOR, and SuperAmp are trademarks of Miltenyi Biotec GmbH.Copyright © 2008 Miltenyi Biotec GmbH. All rights reserved.