Chat with an Expert

MyeloCult™

Media for Long-Term Culture of Human and Mouse Hematopoietic Progenitor Cells

MyeloCult™ media have been developed for the long-term culture of hematopoietic progenitor cells in association with adherent stromal cells. An important application of MyeloCult™ is the quantitation of primitive hematopoietic progenitor cells in the long-term culture-initiating cell (LTC-IC) assay. In this assay hematopoietic cells are cultured in MyeloCult™ medium with supportive human or mouse stromal cell layers and tested for their ability to generate myeloid clonogenic progenitors for several weeks as assayed by replating the cells in a colony-forming unit (CFU) assay using MethoCult™ medium.

Why Use MyeloCult™?

  • Establish stromal cell layers and culture hematopoietic progenitors in “Dexter-type” long-term bone marrow cultures.1,2
  • Perform LTC-IC assays in MyeloCult™ media.3
  • Study regulators of myelopoiesis in long-term cultures.3

MyeloCult™ H5100

Species:

Human

For Initiation and Maintenance of:

  • Myeloid long-term cultures of human hematopoietic cells
  • Stromal cell feeder layers (e.g. M2-10B4 mouse bone marrow fibroblast cell line)

MyeloCult™ M5300

Species:

Mouse

For Initiation and Maintenance of:

  • Myeloid long-term cultures of mouse hematopoietic cells
  • Stromal cell feeder layers (e.g. M2-10B4 mouse bone marrow fibroblast cell line)
Expand Details
Collapse Details

Protocol

Figure 1. Procedure for Limiting-Dilution Long Term Culture-Initiating Cell (LTC-IC) Assay



Figure 2. Procedure for Bulk Culture LTC-IC Assay



Key Applications

Human Hematopoietic Cell Culture

Nemeth M et al. (2009) beta-Catenin expression in the bone marrow microenvironment is required for long-term maintenance of primitive hematopoietic cells. Stem Cells 27(5): 1109-1119
Tipping A et al. (2009) High GATA-2 expression inhibits human hematopoietic stem and progenitor cell function via effects on cell cycle. Blood
Peterson M et al. (2008) Inhibitory receptor signaling via tyrosine phosphorylation of the adaptor Crk. Immunity 29(4): 578-588
Carlo-Stella C et al. (2007) Placental growth factor-1 potentiates hematopoietic progenitor cell mobilization induced by granulocyte colony-stimulating factor in mice and nonhuman primates. Stem Cells 25(1): 252-261
Pastos K et al. (2006) Differential effects of recombinant thrombopoietin and bone marrow stromal-conditioned media on neonatal versus adult megakaryocytes. Blood 108(10): 3360-3362
Sloand E et al. (2006) Granulocyte colony-stimulating factor preferentially stimulates proliferation of monosomy 7 cells bearing the isoform IV receptor. Proc Natl Acad Sci 103(39): 14483-14488
Pirson L et al. (2006) 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
Takeda A et al. (2006) NUP98-HOXA9 induces long-term proliferation and blocks differentiation of primary human CD34+ hematopoietic cells. Cancer Res 66(13): 6628-6637
Giebel B et al. (2006) Primitive human hematopoietic cells give rise to differentially specified daughter cells upon their initial cell division. Blood 107(5): 2146-2152
Srour E et al. (2005) Modulation of in vitro proliferation kinetics and primitive hematopoietic potential of individual human CD34+CD38-/lo cells in G0. Blood 105(8): 3109-3116
Selleri C et al. (2005) Involvement of the urokinase-type plasminogen activator receptor in hematopoietic stem cell mobilization. Blood 105(5): 2198-2205
Giebel B et al. (2004) Segregation of lipid raft markers including CD133 in polarized human hematopoietic stem and progenitor cells. Blood 104(8): 2332-2338
Kogler G et al. (2004) A new human somatic stem cell from placental cord blood with intrinsic pluripotent differentiation potential. J of Exp Med 200(2): 123-135
Guan Y et al. (2003) Detection, isolation, and stimulation of quiescent primitive leukemic progenitor cells from patients with acute myeloid leukemia (AML). Blood 101(8): 3142-3149
Bartolovic K et al. (2004) Inhibitory effect of imatinib on normal progenitor cells in vitro. Blood 103(2): 523-529
Stier S et al. (2003) Ex vivo targeting of p21Cip1/Waf1 permits relative expansion of human hematopoietic stem cells. Blood 102(4): 1260-1266
Danet G et al. (2003) Expansion of human SCID-repopulating cells under hypoxic conditions. J of Clin Invest 112(1): 126-135
Dybedal I et al. (2003) Human reconstituting hematopoietic stem cells up-regulate Fas expression upon active cell cycling but remain resistant to Fas-induced suppression. Blood 102(1): 118-126
Punzel M et al. (2003) The symmetry of initial divisions of human hematopoietic progenitors is altered only by the cellular microenvironment. Exp Hematol 31(4): 339-347
Murdoch B et al. (2003) Wnt-5A augments repopulating capacity and primitive hematopoietic development of human blood stem cells in vivo. Proc Natl Acad Sci 100(6): 3422-3427
Kootstra N et al. (2003) Abrogation of postentry restriction of HIV-1-based lentiviral vector transduction in simian cells. Proc Natl Acad Sci U S A 100(3): 1298-1303
Bhatia M et al. (1998) A newly discovered class of human hematopoietic cells with SCID-repopulating activity. Nat Med 4(9): 1038-1045
Dybedal I et al. (2003) Human reconstituting hematopoietic stem cells up-regulate Fas expression upon active cell cycling but remain resistant to Fas-induced suppression. Blood 102(1): 118-126

Mouse Hematopoietic Cell Culture

Zimmer S et al. (2011) Crebbp haploinsufficiency in mice alters the bone marrow microenvironment leading to loss of stem cells and excessive myelopoiesis. Blood
Lymperi S et al. (2010) Inhibition of osteoclast function reduces hematopoietic stem cell numbers in vivo. Blood 117(5): 1540-1549
Ohno Y et al. (2010) Hoxb4 transduction down-regulates Geminin protein, providing hematopoietic stem and progenitor cells with proliferation potential. Proc Natl Acad Sci
Ohmori T et al. (2010) Vinculin is indispensable for repopulation by hematopoietic stem cells, independent of integrin function.J Biol Chem 285(41): 31763-31773
Zhang Q et al. (2010) Fancd2-/- mice have hematopoietic defects that can be partially corrected by resveratrol. Blood
Sumitomo A et al. (2010) Transcriptional Mediator Subunit MED1/TRAP220 in Stromal Cells Is Involved in Hematopoietic Stem/Progenitor Cell Support through Osteopontin Expression. Mol Cell Biol
Pandit T et al. (2006) Sustained exposure to nicotine leads to extramedullary hematopoiesis in the spleen.Stem cells 24(11): 2373-2381
Lee Y et al. (2006) CCAAT/enhancer binding proteins alpha and epsilon cooperate with all-trans retinoic acid in therapy but differ in their antileukemic activities. Blood 108(7): 2416-2419
Carella C et al. (2006) The ETS factor TEL2 is a hematopoietic oncoprotein. Blood 107(3): 1124-1132
Carmen M et al. (2004) Interleukin-6 deficiency affects bone marrow stromal precursors, resulting in defective hematopoietic support.Blood 103(9): 3349-3354
Lacout C et al. (2003) A defect in hematopoietic stem cell migration explains the nonrandom X-chromosome inactivation in carriers of Wiskott-Aldrich syndrome. Blood 102(4): 1282-1289
Truong B et al. (2003) CCAAT/Enhancer binding proteins repress the leukemic phenotype of acute myeloid leukemia. Blood 101(3): 1141-1148
Jin H et al. (2002) Intracerebral transplantation of mesenchymal stem cells into acid sphingomyelinase-deficient mice delays the onset of neurological abnormalities and extends their life span. J of Clin Invest 109(9): 1183-1191
Samper E et al. (2002) Long-term repopulating ability of telomerase-deficient murine hematopoietic stem cells.Blood 99(8): 2767-2775
Iversen P et al. (2002) Decreased hematopoiesis in bone marrow of mice with congestive heart failure. Am J of Physiol, Regul, Integr Comp Physiol 282(1): R166-72
Jasinski M et al. (2001) GATA1-Cre mediates Piga gene inactivation in the erythroid/megakaryocytic lineage and leads to circulating red cells with a partial deficiency in glycosyl phosphatidylinositol-linked proteins (paroxysmal nocturnal hemoglobinuria type II cells). Blood 98(7): 2248-2255

References

  1. Dexter T et al. (1977) Conditions controlling the proliferation of haematopoietic stem cells in vitro. J Cell Physiol 91(3): 335–44
  2. Gartner S et al. (1980) Long-term culture of human bone marrow cells. Proc Natl Acad Sci 77(8): 4756–9
  3. Sutherland H et al. (1990) Functional characterization of individual human hematopoietic stem cells cultured at limiting dilution on supportive marrow stromal layers. Proc Natl Acad Sci 87(9): 3584–8
Copyright © 2018 by STEMCELL Technologies Inc. All rights reserved including graphics and images. STEMCELL Technologies & Design, STEMCELL Shield Design, Scientists Helping Scientists, AGGREWELL, CELLADHERE, CLONACELL, EASYPLATE, EASYSEP, ENDOCULT, EPICULT, ESCULT, ERYTHROCLEAR, FRESR, MAMMOCULT, MEGACULT, MESENCULT, METHOCULT, MYELOCULT, NEUROCULT, PNEUMACULT, PROSTACULT, RELESR, ROBOSEP, ROSETTESEP, SEPMATE, SMARTDISH, SPINSEP, STEMDIFF, STEMSEP, STEMSPAN, and STEMVISION are trademarks of STEMCELL Technologies Canada Inc. All other trademarks and registered trademarks are the property of their respective holders. While STEMCELL has made all reasonable efforts to ensure that the information provided by STEMCELL and its suppliers is correct, it makes no warranties or representations as to the accuracy or completeness of such information.