tissue regener research br

MACS® Products for tissue regeneration research 3

MACS Technology—the complete solution 4

MACS Cell Separation Strategies 5

Human stem cells—sample preparation, cell separation and analysis 6

Embryonic stem cells and induced pluripotent stem cells—separation and analysis 7

Mesenchymal stromal cells—separation and analysis 8

Endothelial progenitor cells—separation and analysis 9

Tissue regeneration—examples 10

References 12

MACS Products for sample preparation, cell separation, and cell analysis 13

MACS Products for cell culture 14

Molecular biology products and services 15

Cover image: SPL/Agentur Focus

Table of contents

MACS® Products for tissue regeneration research

MACS® Products provide the basis for reliable methods to investigate the full potential of stem cells in tissue regeneration research.Stem and progenitor cells have the potential to revolutionize

tissue regeneration and engineering in the future. The

understanding of stem cell biology is of paramount importance

for the development of stem cell–based therapies. The

multipotent—and even pluripotent—differentiation capacity

of certain stem and progenitor cell populations makes them a

potential key to successful regeneration of many tissue types.

Over the coming decade, research into this field may take center

stage as the new frontier in the treatment of many disabling

diseases and injuries.

Reproducible data, standardized protocols, and defined

cell sources are key to unravel the full potential of stem and

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.

MACS® Technology

3

MACS Products provide the tools that open up new

perspectives for tissue regeneration research:

• Reliable isolation of defined stem cell populations

with excellent purity for reproducible results

• Stem cells and progenitors thereof can be isolated

within an hour rather than several days,

saving valuable time for you to focus on your research.

• Isolated cells can immediately be used for cell culture

and animal experiments; cells remain viable and fully

functional.

• Miltenyi Biotec offers integrated solutions for a wide

variety of research needs—from sample preparation

to molecular analysis.

MACS® Technology —the complete solution

MACS® Technology

4

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.

Magnetic labeling

Cells of interest are labeled with MACS® MicroBeads in a short incubation step.

Magnetic separation

Labeled and unlabeled cells are separated on a MACS Column placed in the magnetic field of a MACS Separator. The flow-through can be collected as the non-magnetic, unlabeled cell fraction.

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.

MACS Technology

MACS® Technology—the gold standard in cell separationMACS Technology is based on MACS MicroBeads, MACS

Columns, and MACS Separators—strong permanent magnets.

MicroBeads are superparamagnetic particles coupled to specific

monoclonal antibodies.

Target cells can be magnetically isolated by positive selection

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 greater

flexibility for the sequential sorting of complex subpopulations

of cells.

MACS MicroBeads

• Highly specific monoclonal antibody conjugates

• Small (50 nm), virus-sized nanoparticles

• Non-toxic, biodegradable

• Colloidal, for ease of handling and short incubation times

MACS Columns and MACS Separators

• Optimal recovery and high purity with MACS Columns

• Gentle to cells

• Automated cell separation with

autoMACS™ Pro Separator

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.)

Benefits of MACS® Technology at a glance:

• Fast—cell separation takes less than one hour

• Gentle—separated cells remain viable and functional

• Flexible—both labeled and unlabeled fractions can be

obtained with excellent purity and high recovery

• Easy separation of large cell numbers—up to 109

labeled cells per column

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.

5

MACS® Cell Separation Strategies

Sample preparationFor 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™ DissociatorThe 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

• Standardized procedure

• Reliable and reproducible results

• High level of user safety

• Sterile sample handling

Cell separation and analysisFor 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.

Human stem cells in tissue regeneration researchStem 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.

The gentleMACS Dissociator—the gentle way of automated tissue dissociation

6

Human stem cells—sample preparation, cell separation and analysis

Sample preparation, cell separation, cell analysis

Embryonic stem cells and induced pluripotent stem cellsPluripotent 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

in the future. For these applications it is crucial to use homogenous cell populations. MACS®

Technology enables the isolation of numerous particular cell types and subsets at high

purity—ready for in vitro and animal experiments.

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.

After their in vitro differentiation, ESC- or iPS-derived cells exhibit a broad cellular

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

Cell separation

Cell analysis

microRNA expression profiling

Gene expression analysis

For more information on Miltenyi Biotec products for ESC and iPS cell research, please refer to the corresponding brochure that is available for download at www.miltenyibiotec.com.

7

Cell separation and analysis

Embryonic stem cells and induced pluripotent stem cells— separation and analysis

Mesenchymal stromal cellsMesenchymal 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 diff erentiate 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.

Bone marrowCD271 is a well-known marker for the isolation of MSCs from bone marrow.20–22

Marrow stromal antigen-1 (MSCA-1), which is recognized by clone W8B2, was shown to be

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 identifi cation of MSCs with a high

proliferative potential.

Additional markers for the isolation of MSCs from bone marrow include CD117 23, CD105 24,25,

Stro-1 26 , CD146 27, and CD13328.

Multipotent adult progenitor cells (MAPCs) can be enriched from bone marrow by

depletion of CD235a (glycophorin A) and CD45.33

Cord bloodCD133 is a suitable marker for the identifi cation of MSCs from cord blood.17

LipoaspirateVarious strategies can be pursued for the isolation of MSCs from lipoaspirate: positive

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 pulpSTRO-1 and CD146 are appropriate markers for the separation of MSCs from dental pulp.27

Amniocentesis culturesMSC-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 diff erentiate into cell types that represent

all embryonic germ layers. 32

Sample preparation

Cell separation

Cell analysis

Cell culture

Expression profi ling

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.

BM-MNCs before separation

MSCA-1+ cells

Anti-MSCA-1 (W8B2)-APC

CD

45-F

ITC

Anti-MSCA-1 (W8B2)-APC

CD

45-F

ITC

For more information on Miltenyi Biotec products for MSC research, please refer to the corresponding brochure available for download at www.miltenyibiotec.com.

8

Mesenchymal stromal cells—separation and analysis

Cell 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 vascular

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.

SPECT (single-photon emission computed tomographic) scan from the posterior wall area of a human heart. Courtesy of Prof. Gustav Steinhoff, Rostock, Germany.

9

Endothelial progenitor cells—separation and analysis

Tissue-resident stem cells have been found in almost any tissue.

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 tissueIn 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

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

muscle tissue51 and skeletal muscle52, respectively. Cardiac

muscle regeneration is a prime goal in cardiovascular disease

research; functional cardiomyocytes can be generated in vitro

from ESCs15, CD133+ fetal liver cells53, and from CD34+ cEPCs from

peripheral blood54.

Tissue regeneration research—examples

CD133+ cells, isolated from mobilized peripheral blood, gave rise to adherent cells after 3–5 weeks of cultivation. These cells were able to differentiate into hepatocyte-like cells. (A) The cells are stained for hepatocyte nuclear factor-3 (FITC), albumin (Cy3), and nuclei (DAPI). (B) The cells are stained for cytokeratin 19 (Cy3) and nuclei (DAPI) (200×). (Courtesy of Selim Kuçi, Tübingen, Germany.)

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.)

A

B

10

Tissue regeneration research

Neural tissueThe 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 diff erentiate

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 diff erentiated 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

Tissue regeneration research—examples

B

A

C

CD133+ 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 Biotec products for neuroscience research, please refer to the corresponding brochure that is available for download at www.miltenyibiotec.com.

11

MACS® Technology

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.

References

12

References

13

MACS® Products

MACS® Products for sample preparation, cell separation, and cell analysis

Product Components or capacity Order no.

gentleMACS™ Starting Kit

gentleMACS™ Dissociator C Tubes, 25 pieces M Tubes , 25 piecesPower cord User manual gentleMACS Protocols

130-093-235

C Tubes C Tubes, 25 pieces, single-packed

130-093-237

M Tubes M Tubes, 25 pieces, single-packed M Tubes, 50 pieces per bag

130-093-236 130-093-458

Neural Tissue Dissociation Kit (P)

50 tests with up to 400 mg of neural tissue

130-092-628

Neural Tissue Dissociation Kit (T)

50 tests with up to 400 mg of neural tissue

130-093-231

Pre-Separation Filters

50 filters 130-041-407

Dead Cell Removal Kit

For 10⁹ total cells 130-090-101

Product Order no.

Anti-A2B5 MicroBeads, human, mouse, rat 130-093-388

MSC Research Tool Box – MSCA-1 (W8B2), human

130-093-572

Anti-MSCA-1 (W8B2) MicroBead Kit, human 130-093-583

Anti-PSA-NCAM MicroBeads, human, mouse, rat

130-092-966

CD31 MicroBead Kit, human 130-091-935


CD34 MultiSort Kit, human 130-056-701

CD45 MicroBeads, human 130-045-801






CD235a (Glycophorin A) MicroBeads, human 130-050-501

MSC Research Tool Box – CD271 (PE), human 130-092-867

MSC Research Tool Box – CD271 (APC), human 130-092-291

CD271 MicroBead Kit (PE), human 130-092-819

Lineage Cell Depletion Kit, human 130-092-211

CD271 MicroBead Kit (APC), human 130-092-283

CD326 (EpCAM) MicroBeads, human 130-061-101

MACS Products for the analysis of stem and progenitor cells

Product Order no.

Anti-A2B5-PE, -APC, pure, human, mouse, rat

130-093-581, 130-093-582, 130-092-394

Anti-MSCA-1 (W8B2)-FITC, -PE, -APC, -Biotin, pure, human

130-093-585, 130-093-587, 130-093-589, 130-093-593, 130-093-595

Anti-PSA-NCAM-PE, -APC, human, mouse, rat

130-093-274, 130-093-273

CD31-FITC, -PE, -APC, human 130-092-64, 130-092-653, 130-092-652

CD34-FITC, -PE, -APC, human 130-081-001, 130-081-002, 130-090-954

CD56-PE, APC, pure, human 130-090-755, 130-090-843, 130-090-955

CD117 (A3C6E2)-PE, -APC, human 130-091-734, 130-091-733

CD117 (AC126)-PE, human 130-091-735

CD133/1 (AC133)-PE, -APC, -Biotin, pure, human

130-080-801, 130-090-826, 130-090-664, 130-090-422

CD133/2 (293C3)-PE, -APC, -Biotin, pure, human

130-090-853, 130-090-854, 130-090-852, 130-090-851

CD133/1 (W6B3C1) pure, human 130-092-395

CD146-FITC, -PE, -APC, -Biotin, pure, human

130-092-851, 130-092-853, 130-092-849, 130-092-852, 130-092-850

CD271 (LNGFR), -PE, -APC, -Biotin, human

130-091-885, 130-091-884, 130-091-883

CD309 (VEGFR/KDR)-PE, -APC, -Biotin, human

Coming soon

CD326 (EpCAM)-FITC, -PE, -APC, human

130-080-301, 130-091-253, 130-091-254

EPC Enrichment and Enumeration Kit, human

130-093-477

MACS Products for sample preparation

Product Order no.

Anti-FITC MicroBeads 130-048-701

Anti-PE MicroBeads 130-048-801

Anti-APC MicroBeads 130-090-855

Anti-Cy5/Anti-Alexa Fluor 647 MicroBeads 130-091-395

Anti-Cy7 MicroBeads 130-091-652

Anti-Biotin MicroBeads 130-090-485

Streptavidin MicroBeads 130-048-101

Rat Anti-Mouse IgG1 MicroBeads 130-047-101

Rat Anti-Mouse IgG2a+b MicroBeads 130-047-201

Rat Anti-Mouse IgM MicroBeads 130-047-301

Goat Anti-Mouse IgG MicroBeads 130-048-401

Mouse Anti-Rat Kappa MicroBeads 130-047-401

Goat Anti-Rat IgG MicroBeads 130-048-501

Goat Anti-Rabbit IgG MicroBeads 130-048-602

MACS Products for indirect magnetic labeling

MACS Products for the isolation of stem and progenitor cells

MACS® Technology

MACS® Products for cell culture

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NH media for the enumeration, expansion, and differentiation of MSCsMSCs 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 tissue or organ within the human body. Miltenyi Biotec also 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.

AdipocytesNH AdipoDiff Medium

ChondrocytesNH ChondroDiff Medium

OsteoblastsNH OsteoDiff Medium

MACS® NH Media enable the differentiation of mesenchymal stromal cells (MSCs) into functional nonhematopoietic (NH) cell types.

MSC expansionNH Expansion Medium

MSC enumerationNH CFU-F Medium

NH stem cell source, e.g., bone marrow, lipoaspirate

Basic media and cytokines for stem cell cultureMiltenyi Biotec offers cell culture media for a broad spectrum of applications. In addition, high-quality recombinant cytokines and growth factors are available, that are well-suited for various applications such as cell culture, differentiation studies, and biological assays. Selected products are available in a premium-grade format with excellent purity and high, well-defined activity as well as in research-grade quality. For a complete list of available cytokines, please visit http://www.miltenyibiotec.com/cytokines.

CytoMix – MSC, humanThe CytoMix – MSC, human (130-093-552) is a composition of 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 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

MACS Basic Culture Media Order no.

DMEM 500 mL 130-091-437

DMEM with stable glutamine 500 mL 130-091-438

RPMI 1640 500 mL 130-091-440

RPMI 1640 with stable glutamine 500 mL 130-091-439

MACS Cytokines and Growth Factors

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; 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)

MACS® Products

Molecular biology products and services

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Stem cell differentiation tracking by gene expression profilingMACSmolecular 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 from 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 from just a few to as many as 10⁷ cells.

PIQOR™ Stem Cell MicroarrayThe PIQOR™ Stem Cell Microarray comprises 942 relevant marker genes for human stem cells and their differentiation. It is available as a convenient microarray kit* or within the scope of the microarray service**. Gene expression experiments allow for the quality control of different stem cell types, comparison between different stages of differentiation, as well as the optimization of differentiation protocols.

SuperAmp™ Service ***When the number of stem cells for analysis is low, Miltenyi Biotec offers the ideal solution for gene expression profiling needs. The SuperAmp™ Service (available as an extension of the PIQOR™ Microarray Service) can reliably amplify mRNA million-fold from as little as one cell. The service is ideal for stem cells sorted with MACS® Technology, flow cytometry, or even from laser capture microdissected tissue.

miRXplore™ Kits and ServicesExplore microRNA expression in human and mouse stem cells with the new miRXplore™ Microarray Kits and Services. Designed in collaboration with experts at the Rockefeller University¹, the microarray covers more than 2700 human, mouse, rat, and viral microRNA sequences and possess rigorous internal control system. Sequences differing by just one oligonucleotide can be reproducibly detected and re-ratios calculated with the use of the proprietary miRXplore Universal Reference.

Reference1. Landgraf, P. et al. (2007) Cell 129: 1401–1414.

a-Hyb™ Hybridization Station

* PIQOR™ Microarray Kits are not available in the US and Canada.** 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 also available.

*** In combination with the Microarray Services only. The SuperAmp Service is not available for microRNA amplification

mRNA isolation/cDNA synthesis

µMACS mRNA Isolation Kit Small ScaleLarge ScaleFor Total RNA

130-075-201130-075-101130-075-102

µMACS mRNA Isolation Starting Kit 130-075-202

µMACS One-step cDNA Kit 130-091-902

µMACS One-step cDNA Starting Kit 130-091-989

PIQOR Microarray Kit *

PIQOR Stem Cell Microarray Kit, antisense

4 Microarrays 130-092-033


PIQOR Microarray Service **

Service Stem Cell Microarray Plus Amplification 160-000-765

SuperAmp Amplification ***

SuperAmp Service (per sample) 160-000-936

miRXplore Microarray Kit



miRXplore Microarray Services

miRXplore Microarray Service 160-001-143

miRXplore Universal Reference Service 160-001-161

miRXplore Additional Total RNA Extraction 160-001-162

Miltenyi BiotecMiltenyi Biotec was founded in 1989; nowadays, more than 1100 employees develop, produce, and sell innovative products for cell research and clinical applications. Due to an intense focus on stem cell research at Miltenyi Biotec, the portfolio includes the largest range of products available for the separation of stem cells. Moreover, Miltenyi Biotec offers integrated solutions for a wide variety of research needs—from sample preparation to molecular analysis.

Miltenyi Biotec Asia Pacific Pte. Ltd. (Singapore)Phone +65 6238 [email protected]

Miltenyi Biotec S.L. (Spain)Phone +34 91 512 12 [email protected]

Miltenyi Biotec Ltd. (UK)Phone +44 1483 799 800 [email protected]

Miltenyi Biotec SAS (France)Phone +33 1 56 98 16 [email protected]

Miltenyi Biotec S.r.l. (Italy)Phone +39 051 646 [email protected]

Miltenyi Biotec K.K. (Japan)Phone +81 3 5646 [email protected]

Miltenyi Biotec B.V. (Benelux)[email protected] service NetherlandsPhone 0800 4020120Customer service BelgiumPhone 0800 94016Customer service LuxembourgPhone 800 24971

Miltenyi Biotec Trading (Shanghai) Co., Ltd. (P.R. China) Phone +86 21 6235 [email protected]

Miltenyi Biotec Inc. 12740 Earhart AvenueAuburn, CA 95602, USAPhone 800 FOR MACS, +1 530 888 8871 Fax +1 530 888 [email protected]

Miltenyi Biotec Australia Pty. Ltd.Phone +61 02 8877 [email protected]

Miltenyi Biotec GmbH Friedrich-Ebert-Straße 6851429 Bergisch Gladbach GermanyPhone +49 2204 8306-0Fax +49 2204 [email protected]

www.miltenyibiotec.com

Unless 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.

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