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MegaCult™

Collagen-Based Medium for the Detection of Human and Mouse 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:

Human

Cytokines:

Available with or without recombinant cytokines

Contains:

  • Collagen solution
  • Medium
  • CFU-Mk staining kit
  • Double chamber slides

Applications:

Culture and enumeration of megakaryocytic progenitors in CFU-Mk assays on human cord blood, bone marrow and peripheral blood.

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:

Available with added recombinant cytokines (human only) or without (human or mouse)

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):

50 mL bottle

Species:

  • Human
  • Mouse

Cytokines:

No added 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.
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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

Pessina A et al. (2009) Application of human CFU-Mk assay to predict potential thrombocytotoxicity of drugs. Toxicol In Vitro 23(1): 194-200
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