²Ñ±ð²µ²¹°ä³Ü±ô³Ùâ„¢ - Colony Assays of Megakaryocyte Progenitors (CFU-Mk)
MegaCult™ Media for CFU-Mk Assays
Collagen-Based Medium for Colony Assays of Megakaryocyte Progenitors
MegaCult™-C is a collagen-based culture system for colony-forming unit (CFU) assays of megakaryocyte progenitors (CFU-Mk) from human or mouse hematopoietic tissues.1 Megakaryocyte colonies are difficult to detect by morphological criteria alone as colonies may contain as few as three cells and megakaryocytes cannot be easily distinguished morphologically from other cell lineages, particularly macrophages. Unlike methylcellulose-based media used in CFU assays for other lineages, collagen gels used in MegaCult™ can be readily dehydrated for subsequent cytochemical or immunocytochemical staining. Human CFU-Mk colony detection in MegaCult™ is based on immunocytochemical staining for the megakaryocyte marker CD41, while mouse colony detection is based on acetylcholinesterase activity.
MegaCult™ medium is available in formats with or without cytokines or added lipids. MegaCult™ media with cytokines contain recombinant human (rh) IL-3, IL-6 and TPO for optimal growth of human CFU-Mk. MegaCult™ media without cytokines is recommended for applications where control over the types and amounts of cytokines to use is important, for example for investigation of factors influencing the growth and maturation of megakaryocytic progenitor cells. MegaCult™-C media without cytokines is also used for the culture and enumeration of mouse CFU-Mk after addition of appropriate cytokines, such as recombinant mouse IL-3, rhIL-6, rhIL-11, rhTPO and/or other factors, chosen by the user.
MegaCult™-C
Complete Kits
Format (Media):
- 24 x 2 mL tubes (with cytokines)
- 24 x 1.7 mL tubes (medium only)
Species:
Cytokines:
Contains:
- Collagen solution
- Medium
- CFU-Mk staining kit
- Double chamber slides
Applications:
MegaCult™-C
Collagen and Medium
Format (Media):
- 24 x 2 mL tubes (with cytokines)
- 24 x 1.7 mL tubes (medium only)
Species:
- Human
- Mouse
Cytokines:
Contains:
- Collagen solution
- Medium
Applications:
- CFU-Mk assays on human cord blood, bone marrow and peripheral blood.
- CFU-Mk assays on mouse bone marrow. The addition of cytokines is required.
MegaCult™-C
Collagen and Medium with Lipids
Format (Media):
Species:
- Human
- Mouse
Cytokines:
Contains:
- Collagen solution
- Medium with lipids
Applications:
- CFU-Mk assays on progenitor-enriched preparations of human cord blood, bone marrow and peripheral blood. The addition of cytokines is required.
- CFU-Mk assays on mouse bone marrow. The addition of cytokines is required.
- CFU assays for other progenitor cells after addition of appropriate cytokines chosen by the user.
Data
Figure 1. Examples of Small and Medium Sized CFU-Mk Colonies Derived From Human Megakaryocyte Progenitors
Protocol
Figure 2. Procedure for Assays of Human Megakaryocyte Progenitors
Figure 3. Procedure for Assays of Mouse Megakaryocyte Progenitors
Key Applications
Drug Toxicity Testing
Transplantation and Engraftment Kinetics of Human HSPCs
Pirson L et al. (2006) Stem Cells 24(7): 1814-1821
Megakaryocyte Biology
Boyer L et al. (2008) J Immunol Methods 332(1-2): 82-91
Vodyanik M et al. (2006) Blood 108(6): 2095-2105821
HSPC Biology
Migliaccio A et al. (2003) J Exp Med 197(3): 281-296
Ito C et al. (2003)
Blood 101(2): 517-523
Wang Y et al. (2011) Genes Dev 25(13): 1426-1438
Lannutti B et al. (2008) Blood 113(8): 1778-1785
Hisa T et al. (2004) EMBO J 23(2): 450-459
Xing S et al. (2008) Blood 111(10): 5109-5117
Wernig G et al. (2006) Blood 107(11): 4274-4281
Moody J et al. (2004) Blood 103(12): 4503-4510
References
- Hogge D et al. (1997) Br J Hematol. 96(4): 790-800.
