MyeloCult® M5300

Recommended for:	Initiation and maintenance of myeloid long-term cultures of mouse hematopoietic cells and stromal feeder layers
Accessory Products:	• Hydrocortisone (Catalog #07904) • 35 mm Treated Tissue Culture Dishes (Catalog #27114/27116) • 96-well Treated Tissue Culture Dishes (Catalog #27135/27136) • Hanks' Balanced Salt Solution Modified (without Ca++ and Mg++) (Catalog #37250) • Minimum Essential Medium Eagle (MEM) Alpha Modification (Catalog #36450) • Trypsin in Citrate Saline (Catalog #07400) • Trypsin-EDTA (Catalog #07901) • Iscove's Modified Dulbecco's Medium (IMDM) with 2% Fetal Bovine Serum (Catalog #07700) • MethoCult® GF H3434 (Catalog #03434/03444)
Intended Use Statement:	For Research Use Only. Not for Therapeutic or Diagnostic Use.
Contains:	• Horse Serum • Fetal Bovine Serum • 2-Mercaptoethanol • MEM Alpha • Supplements
Product Type:	Specialized cell culture media
Application:	Characterization, Colony assays & quantification
Area of Interest:	Cancer, Hematologic malignancies, Hematopoietic stem cell research, Stem cell biology
Cell Type:	Hematopoietic stem & progenitor cells
Medium Type:	Liquid
Popular Product Line:	MyeloCult
Species:	Mouse

Procedures and instruction manuals:

•  MANUAL: Mouse Long-Term Culture-Initiating Cell (LTC-IC) Assays Procedure Manual
•  PRODUCT INFORMATION SHEET - 05300, Lot# All:29560-PIS_1_2_0.pdf
•  PRODUCT INFORMATION SHEET - 05350, Lot# All:29560-PIS_1_2_0.pdf

Educational resources:

•  BROCHURE: The World Leader in Tools For Hematopoietic Stem Cell Research
•  BROCHURE: MegaCult™-C and MyeloCult™ For Megakaryocyte Progenitors and Primitive Hematopoietic Progenitors

MSDS:

•  05300_05350-MSDS.pdf

This product has been used in:

1. Yoshinori Ohno et al. Hoxb4 transduction down-regulates Geminin protein, providing hematopoietic stem and progenitor cells with proliferation potential.Proc Natl Acad Sci U S A (November 22, 2010)
2. Tsukasa Ohmori et al. Vinculin is indispensable for repopulation by hematopoietic stem cells, independent of integrin function.J Biol Chem 285 (41) 31763-31773 (October 8, 2010)
3. Qing-Shuo Zhang et al. Fancd2-/- mice have hematopoietic defects that can be partially corrected by resveratrol.Blood (September 8, 2010)
4. Akiko Sumitomo et al. Transcriptional Mediator Subunit MED1/TRAP220 in Stromal Cells Is Involved in Hematopoietic Stem/Progenitor Cell Support through Osteopontin Expression.Mol Cell Biol (August 16, 2010)
5. T Pandit et al. Sustained exposure to nicotine leads to extramedullary hematopoiesis in the spleen.Stem cells (Dayton, Ohio) 24 (11) 2373-2381 (November 2006)
6. Y Lee et al. 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 (October 1, 2006)
7. C Carella et al. The ETS factor TEL2 is a hematopoietic oncoprotein.Blood 107 (3) 1124-1132 (February 1, 2006)
8. G Kogler et al. A new human somatic stem cell from placental cord blood with intrinsic pluripotent differentiation potential.The Journal of experimental medicine. 200 (2) 123-135 (July 19, 2004)
9. C Lacout et al. A defect in hematopoietic stem cell migration explains the nonrandom X-chromosome inactivation in carriers of Wiskott-Aldrich syndrome.Blood 102 (4) 1282-1289 (August 15, 2003)
10. I Dybedal et al. 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 (July 1, 2003)
11. B-T H Truong et al. CCAAT/Enhancer binding proteins repress the leukemic phenotype of acute myeloid leukemia.Blood 101 (3) 1141-1148 (February 1, 2003)
12. H Jin et al. Intracerebral transplantation of mesenchymal stem cells into acid sphingomyelinase-deficient mice delays the onset of neurological abnormalities and extends their life span.The Journal of clinical investigation 109 (9) 1183-1191 (May 2002)
13. E Samper et al. Long-term repopulating ability of telomerase-deficient murine hematopoietic stem cells.Blood 99 (8) 2767-2775 (April 15, 2002)
14. P Iversen et al. Decreased hematopoiesis in bone marrow of mice with congestive heart failure.American journal of physiology. Regulatory, integrative and comparative physiology 282 (1) R166-72 (January 2002)
15. M Jasinski et al. 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 (October 1, 2001)

Background References:

1. G Kogler et al. An eight-fold ex vivo expansion of long-term culture-initiating cells from umbilical cord blood in stirred suspension cultures.Bone marrow transplantation 21 Suppl 3 S48-53 (June 1998)
2. F Bertolini et al. Multilineage long-term engraftment potential of drug-resistant hematopoietic progenitors.Blood 90 (8) 3027-3036 (October 15, 1997)
3. L E Ailles et al. Detection and characterization of primitive malignant and normal progenitors in patients with acute myelogenous leukemia using long-term coculture with supportive feeder layers and cytokines.Blood 90 (7) 2555-2564 (October 1, 1997)
4. F Prosper et al. Primitive long-term culture initiating cells (LTC-ICs) in granulocyte colony-stimulating factor mobilized peripheral blood progenitor cells have similar potential for ex vivo expansion as primitive LTC-ICs in steady state bone marrow.Blood 89 (11) 3991-3997 (June 1, 1997)
5. S Ghaffari et al. Diverse effects of anti-CD44 antibodies on the stromal cell-mediated support of normal but not leukaemic (CML) haemopoiesis in vitro.Br J Haematol 97 (1) 22-28 (April 1997)
6. P W Zandstra et al. Cytokine manipulation of primitive human hematopoietic cell self-renewal.Proc Natl Acad Sci U S A 94 (9) 4698-4703 (April 29, 1997)
7. H Schrezenmeier et al. Quantitative analysis of cobblestone area-forming cells in bone marrow of patients with aplastic anemia by limiting dilution assay.Blood 88 (12) 4474-4480 (December 15, 1996)
8. M Cavazzana-Calvo et al. Role of interleukin-2 (IL-2), IL-7, and IL-15 in natural killer cell differentiation from cord blood hematopoietic progenitor cells and from gamma c transduced severe combined immunodeficiency X1 bone marrow cells.Blood 88 (10) 3901-3909 (November 15, 1996)
9. F Bertolini et al. A new method for placental/cord blood processing in the collection bag. I. Analysis of factors involved in red blood cell removal.Bone Marrow Transplant 18 (4) 783-786 (October 1996)
10. A L Petzer et al. Characterization of primitive subpopulations of normal and leukemic cells present in the blood of patients with newly diagnosed as well as established chronic myeloid leukemia.Blood 88 (6) 2162-2171 (September 15, 1996)
11. J P Maciejewski et al. A severe and consistent deficit in marrow and circulating primitive hematopoietic cells (long-term culture-initiating cells) in acquired aplastic anemia.Blood 88 (6) 1983-1991 (September 15, 1996)
12. A L Petzer et al. Differential cytokine effects on primitive (CD34+CD38-) human hematopoietic cells: novel responses to Flt3-ligand and thrombopoietin.J Exp Med 183 (6) 2551-2558 (June 1, 1996)
13. L Ponchio et al. Quantitation of the quiescent fraction of long-term culture-initiating cells in normal human blood and marrow and the kinetics of their growth factor-stimulated entry into S-phase in vitro.Blood 86 (9) 3314-3321 (November 1, 1995)
14. M E Lemieux et al. Characterization and purification of a primitive hematopoietic cell type in adult mouse marrow capable of lymphomyeloid differentiation in long-term marrow "switch" cultures.Blood 86 (4) 1339-1347 (August 15, 1995)
15. P W Zandstra et al. Expansion of hematopoietic progenitor cell populations in stirred suspension bioreactors of normal human bone marrow cells.Biotechnology (N Y) 12 (9) 909-914 (September 1994)
16. D A Breems et al. Frequency analysis of human primitive haematopoietic stem cell subsets using a cobblestone area forming cell assay.Leukemia 8 (7) 1095-1104 (July 1994)
17. J H Gong et al. Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells.Leukemia 8 (4) 652-658 (April 1994)
18. C J Eaves et al. Unresponsiveness of primitive chronic myeloid leukemia cells to macrophage inflammatory protein 1 alpha, an inhibitor of primitive normal hematopoietic cells.Proc Natl Acad Sci U S A 90 (24) 12015-12019 (December 15, 1993)
19. C M Verfaillie et al. Soluble factor(s) produced by human bone marrow stroma increase cytokine-induced proliferation and maturation of primitive hematopoietic progenitors while preventing their terminal differentiation.Blood 82 (7) 2045-2053 (October 1, 1993)
20. S Neben et al. Quantitation of murine hematopoietic stem cells in vitro by limiting dilution analysis of cobblestone area formation on a clonal stromal cell line.Exp Hematol 21 (3) 438-443 (March 1993)
21. C Udomsakdi et al. Rapid decline of chronic myeloid leukemic cells in long-term culture due to a defect at the leukemic stem cell level.Proc Natl Acad Sci U S A 89 (13) 6192-6196 (July 1, 1992)
22. H J Sutherland et al. Differential regulation of primitive human hematopoietic cells in long-term cultures maintained on genetically engineered murine stromal cells.Blood 78 (3) 666-672 (August 1, 1991)
23. C J Eaves et al. Mechanisms that regulate the cell cycle status of very primitive hematopoietic cells in long-term human marrow cultures. II. Analysis of positive and negative regulators produced by stromal cells within the adherent layer.Blood 78 (1) 110-117 (July 1, 1991)
24. C C Fraser et al. Expansion in vitro of retrovirally marked totipotent hematopoietic stem cells.Blood 76 (6) 1071-1076 (September 15, 1990)
25. J D Cashman et al. Mechanisms that regulate the cell cycle status of very primitive hematopoietic cells in long-term human marrow cultures. I. Stimulatory role of a variety of mesenchymal cell activators and inhibitory role of TGF-beta.Blood 75 (1) 96-101 (January 1, 1990)
26. H J Sutherland et al. Characterization and partial purification of human marrow cells capable of initiating long-term hematopoiesis in vitro.Blood 74 (5) 1563-1570 (October 1989)
27. S Gartner et al. Long-term culture of human bone marrow cells.Proc Natl Acad Sci U S A 77 (8) 4756-4759 (August 1980)
28. T M Dexter et al. Conditions controlling the proliferation of haemopoietic stem cells in vitro.J Cell Physiol 91 (3) 335-344 (June 1977)
29. A K Stewart et al. In vitro maintenance and retroviral transduction of human myeloma cells in long-term marrow cultures.Cancer Gene Ther 4 (3) 148-156

Product Name	Description	Catalog #
MethoCult® GF M3434	Methylcellulose-Based Medium with Recombinant Cytokines and EPO for Mouse Cells	03434
MyeloCult® H5100	Medium for Human Long-Term Culture of Human Cells	05100
Trypsin in Citrate Solution (0.25%)	0.25% Porcine Trypsin in Citrate Saline Solution	07400
Iscove's MDM with 2% FBS	Medium for Preparing and Washing Samples for Colony-Forming Cell (CFC) Assays	07700
Trypsin-EDTA (0.25%)	Trypsin-EDTA (0.25%)	07901
Hydrocortisone	Hydrocortisone	07904
35 mm Treated Tissue Culture Dishes	35 mm Treated Tissue Culture Dishes	27114
96-Well Treated Tissue Culture Plate	96-Well Treated Tissue Culture Plates	27135
Alpha MEM with Nucleosides	Minimum Essential Medium Eagle - Alpha Modification (Alpha MEM) with Nucleosides	36450
HBSS, Modified (without Ca++ and Mg++)	Hanks' Balanced Salt Solution (HBSS) Without Calcium and Magnesium	37250

Limiting dilution LTC-IC assay

Bulkculture LTC-IC assay