MegaCult®-C Collagen and Medium without Cytokines

Recommended for:	• Assays of human megakaryocyte progenitors in light-density or progenitor-enriched suspensions from bone marrow, cord blood or mobilized peripheral blood • Assays of mouse megakaryocyte progenitors in unseparated or purified cell suspensions of mouse bone marrow
Components:	• MegaCult®-C Medium without Cytokines, 24 x 1.7 mL tubes (Catalog #04900), or • MegaCult®-C Medium without Cytokines, 50 mL (Catalog #04950) • Collagen Solution, 35 mL (Catalog #04902)
Accessory Products:	Double Chamber Slide Kit (includes filter cards and spacers) (Catalog #04963)
Intended Use Statement:	For Research Use Only. Not for Therapeutic or Diagnostic Use.
Product Type:	Specialized cell culture media
Application:	Colony assays & quantification
Area of Interest:	Hematopoietic stem cell research, Stem cell biology
Cell Type:	Hematopoietic stem & progenitor cells
Medium Type:	Collagen-based
Popular Product Line:	MegaCult
Species:	Human, Mouse

Procedures and instruction manuals:

•  MANUAL: MegaCult®-C Assays for Quantitation of Human & Mouse Megakaryocytic Progenitors
•  PRODUCT INFORMATION SHEET - 04960, Lot# All:29553-PIS_2_0_3.pdf
•  PRODUCT INFORMATION SHEET - 04960, Lot# All:29551-PIS_1_0_0.pdf

Educational resources:

•  BROCHURE: The World Leader in Tools For Hematopoietic Stem Cell Research
•  SCIENTIFIC POSTER: Detection of human bipotential erythroid-megakaryocytic progenitors in serum-free collagen gels
•  BROCHURE: MegaCult™-C and MyeloCult™ For Megakaryocyte Progenitors and Primitive Hematopoietic Progenitors

MSDS:

•  04900_04950-MSDS.pdf

FAQS:

• 
  
  • Q. WHY IS THE MEGACULT™-C FORMULATION SERUM FREE?
    A. MegaCult™-C is formulated without FBS to avoid inhibition of CFU-Mk growth by TGF beta and Platelet Factor-4, which are often present in the serum.
  • Q. WHY USE SEMI-SOLID MEDIA?
    A. Semi-solid media (such as methylcellulose-based or collagen-based) allow the clonal progeny of a single progenitor cell to stay together so you can recognize distinct colonies.

This product has been used in:

1. Steven Goossens et al. The EMT regulator Zeb2/Sip1 is essential for murine embryonic hematopoietic stem/progenitor cell differentiation and mobilization.Blood 117 (21) 5620-5630 (May 26, 2011)
2. Fanny Baran-Marszak et al. Expression level and differential JAK2-V617F-binding of the adaptor protein Lnk regulates JAK2-mediated signals in myeloproliferative neoplasms.Blood 116 (26) 5961-5971 (December 23, 2010)
3. Stephanie Halene et al. Serum response factor is an essential transcription factor in megakaryocytic maturation.Blood 116 (11) 1942-1950 (September 16, 2010)
4. Christos Gekas et al. Mef2C is a lineage-restricted target of Scl/Tal1 and regulates megakaryopoiesis and B-cell homeostasis.Blood 113 (15) 3461-3471 (April 9, 2009)
5. Ee-Chun Cheng et al. Role for MKL1 in megakaryocytic maturation.Blood 113 (12) 2826-2834 (March 19, 2009)
6. A Pessina et al. Application of human CFU-Mk assay to predict potential thrombocytotoxicity of drugs.Toxicol In Vitro 23 (1) 194-200 (February 2009)
7. Brian J Lannutti et al. Incomplete restoration of Mpl expression in the mpl-/- mouse produces partial correction of the stem cell repopulating defect and paradoxical thrombocytosis.Blood 113 (8) 1778-1785 (September 16, 2008)
8. Shu Xing et al. Transgenic expression of JAK2V617F causes myeloproliferative disorders in mice.Blood 111 (10) 5109-5117 (May 15, 2008)
9. Lucie Boyer et al. Increased production of megakaryocytes near purity from cord blood CD34+ cells using a short two-phase culture system.J Immunol Methods 332 (1-2) 82-91 (January 18, 2008)
10. Konstantin Levay et al. Tescalcin is an essential factor in megakaryocytic differentiation associated with Ets family gene expression.J Clin Invest 117 (9) 2672-2683 (September 2007)
11. Ting-lei Gu et al. A novel fusion of RBM6 to CSF1R in acute megakaryoblastic leukemia.Blood 110 (1) 323-333 (July 1, 2007)
12. Kuniko Matsumura-Takeda et al. CD41+/CD45+ cells without acetylcholinesterase activity are immature and a major megakaryocytic population in murine bone marrow.Stem Cells 25 (4) 862-870 (April 2007)
13. Joanna M. Tober et al. The megakaryocyte lineage originates from hemangioblast precursors and is an integral component both of primitive and of definitive hematopoiesis.Blood 109 (4) blood-2006-06-031898-1441 (October 24, 2006)
14. Laurence Pirson et al. Despite inhibition of hematopoietic progenitor cell growth in vitro, the tyrosine kinase inhibitor imatinib does not impair engraftment of human CD133+ cells into NOD/SCIDbeta2mNull mice.Stem Cells 24 (7) 1814-1821 (July 2006)
15. Gerlinde Wernig et al. Expression of Jak2V617F causes a polycythemia vera-like disease with associated myelofibrosis in a murine bone marrow transplant model.Blood 107 (11) 4274-4281 (June 1, 2006)
16. David Campard et al. Multilevel regulation of IL-6R by IL-6-sIL-6R fusion protein according to the primitiveness of peripheral blood-derived CD133+ cells.Stem Cells 24 (5) 1302-1314 (May 2006)
17. Chloe Feral et al. Blocking the alpha 4 integrin-paxillin interaction selectively impairs mononuclear leukocyte recruitment to an inflammatory site.The Journal of clinical investigation 116 (3) 715-723 (March 2006)
18. Jennifer L Moody et al. Anemia, thrombocytopenia, leukocytosis, extramedullary hematopoiesis, and impaired progenitor function in Pten+/-SHIP-/- mice: a novel model of myelodysplasia.Blood 103 (12) 4503-4510 (June 15, 2004)
19. Yuki Kimura et al. Targeted mutations of the juxtamembrane tyrosines in the Kit receptor tyrosine kinase selectively affect multiple cell lineages.Proc Natl Acad Sci U S A 101 (16) 6015-6020 (April 20, 2004)
20. Qi Wang et al. BUBR1 deficiency results in abnormal megakaryopoiesis.Blood 103 (4) 1278-1285 (February 15, 2004)
21. Tomoyuki Hisa et al. Hematopoietic, angiogenic and eye defects in Meis1 mutant animals.EMBO J 23 (2) 450-459 (January 28, 2004)
22. Ewout J Houwerzijl et al. Ultrastructural study shows morphologic features of apoptosis and para-apoptosis in megakaryocytes from patients with idiopathic thrombocytopenic purpura.Blood 103 (2) 500-506 (January 15, 2004)
23. Kazuta Yasui et al. Differences between peripheral blood and cord blood in the kinetics of lineage-restricted hematopoietic cells: implications for delayed platelet recovery following cord blood transplantation.Stem Cells 21 (2) 143-151 (2003)
24. Anna Rita Migliaccio et al. GATA-1 as a regulator of mast cell differentiation revealed by the phenotype of the GATA-1low mouse mutant.J Exp Med 197 (3) 281-296 (February 3, 2003)
25. Caryn Y Ito et al. Hematopoietic stem cell and progenitor defects in Sca-1/Ly-6A-null mice.Blood 101 (2) 517-523 (January 15, 2003)
26. I Dobo et al. Comparison of four serum-free, cytokine-free media for analysis of endogenous erythroid colony growth in polycythemia vera and essential thrombocythemia.Hematol J 2 (6) 396-403 (2001)
27. I Dobo et al. Endogenous erythroid and megakaryocytic colony formation in serum-free, cytokine-free collagen gels.J Hematother Stem Cell Res 8 (6) 601-607 (December 1999)
28. I Dobo et al. Collagen matrix: an attractive alternative to agar and methylcellulose for the culture of hematopoietic progenitors in autologous transplantation products.J Hematother 4 (4) 281-287 (August 1995)
29. R Berthier et al. Serum-free medium allows the optimal growth of human megakaryocyte progenitors compared with human plasma supplemented cultures: role of TGF beta.Stem Cells 11 (2) 120-129 (March 1993)

Background References:

1. I Dobo et al. Comparison of four serum-free, cytokine-free media for analysis of endogenous erythroid colony growth in polycythemia vera and essential thrombocythemia.Hematol J 2 (6) 396-403 (2001)
2. I Dobo et al. Endogenous erythroid and megakaryocytic colony formation in serum-free, cytokine-free collagen gels.J Hematother Stem Cell Res 8 (6) 601-607 (December 1999)
3. D Hogge et al. Quantitation and characterization of human megakaryocyte colony-forming cells using a standardized serum-free agarose assay.Br J Haematol 96 (4) 790-800 (March 1997)
4. I Dobo et al. Collagen matrix: an attractive alternative to agar and methylcellulose for the culture of hematopoietic progenitors in autologous transplantation products.J Hematother 4 (4) 281-287 (August 1995)
5. R Berthier et al. Serum-free medium allows the optimal growth of human megakaryocyte progenitors compared with human plasma supplemented cultures: role of TGF beta.Stem Cells 11 (2) 120-129 (March 1993)

Product Name	Description	Catalog #
Double Chamber Slide Kit	For Culture, Fixation and Staining of Collagen Gels in Collagen-Based Assays	04963
MegaCult®-C Staining Kit for CFU-Mk	Staining Kit for Human CFU-Mk	04962
MegaCult®-C Staining Kit for Erythroid Colonies	Staining Kit for Human Erythroid Colonies	04966
IL-3, Human, Recombinant	Interleukin-3	02503
IL-6, Human, Recombinant	Interleukin-6	02506
IL-11, Human, Recombinant	Interleukin-11	02511
TPO, Human, Recombinant	Thrombopoietin	02522
IL-3, Mouse, Recombinant	Interleukin-3	02733
EasySep® Human CD34 Positive Selection Kit	Immunomagnetic Positive Selection Kit	18056
EasySep® Human Whole Blood CD34 Positive Selection Kit	Immunomagnetic Positive Selection Kit	18086
EasySep® Human Cord Blood CD34 Positive Selection Kit	Immunomagnetic Positive Selection Kit	18096
EasySep® Human Progenitor Cell Enrichment Kit	Immunomagnetic Negative Selection Kit	19056
RosetteSep® Human Bone Marrow Progenitor Cell Pre-Enrichment Cocktail	Immunodensity Negative Selection Cocktail	15027
Ficoll-Paque™ PLUS	Density Gradient Medium for the Isolation of Mononuclear Cells	07907
Ficoll-Paque™ PLUS		07917
Ficoll-Paque™ PREMIUM	Density Gradient Medium for the Isolation of Mononuclear Cells	07908
Ficoll-Paque™ PREMIUM		07918
Ammonium Chloride Solution	Reagent for the Lysis of Red Blood Cells	07800
Ammonium Chloride Solution		07850

Procedure summary for assays of human megakaryocytic progenitors

Procedure summary for assays of mouse megakaryocytic progenitors

Examples of colonies derived from human megakaryocyte progenitors