MegaCult™-C Collagen and Medium Without Cytokines

Collagen and medium without cytokines for human and mouse CFU-Mk assays

MegaCult™-C Collagen and Medium Without Cytokines

Collagen and medium without cytokines for human and mouse CFU-Mk assays

MegaCult™-C Collagen and Medium Without Cytokines
1 Kit
1,821 USD
Catalog # 04960

Kit containing collagen and medium without cytokines for human and mouse CFU-Mk assays

MegaCult™-C Medium Without Cytokines
24 x 1.7 mL
1,256 USD
Catalog # 04900

Medium without cytokines

Collagen Solution
35 mL
566 USD
Catalog # 04902

For preparation of collagen gels and for coating cell culture surfaces

What's Included

  • MegaCult™-C Collagen and Medium Without Cytokines (Catalog #04960)
    • Collagen Solution, 35 mL (Catalog #04902)
    • MegaCult™-C Medium Without Cytokines, 24 x 1.7 mL (Catalog #04900)

Overview

MegaCult™-C Collagen and Medium Without Cytokines kit contains medium and collagen solution for colony-forming unit (CFU) assays of human or mouse megakaryocyte progenitor cells (CFU-Mk). The addition of cytokines is required.

MegaCult™-C medium is intended for the culture of CFU-Mk in human bone marrow, mobilized peripheral blood, and cord blood samples, after addition of appropriate cytokines. It is suitable for use with CD34+ enriched cells, mononuclear cells, and cells isolated by other purification methods. It is also intended for assays of megakaryocyte progenitor cells in unseparated or purified cell suspensions of mouse bone marrow, after addition of appropriate cytokines.
Subtype
Semi-Solid Media, Specialized Media
Cell Type
Hematopoietic Stem and Progenitor Cells
Species
Human, Mouse
Application
Cell Culture, Colony Assay, Functional Assay
Brand
MegaCult
Area of Interest
Stem Cell Biology
Formulation
Serum-Free

Data Figures

Procedure Summary for Assays of Human Megakaryocytic Progenitors

Figure 1. Procedure Summary for Assays of Human Megakaryocytic Progenitors

Procedure Summary for Assays of Mouse Megakaryocytic Progenitors

Figure 2. Procedure Summary for Assays of Mouse Megakaryocytic Progenitors

Examples of Colonies Derived From Human Megakaryocyte Progenitors

Figure 3. Examples of Colonies Derived From Human Megakaryocyte Progenitors

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
04960
Lot #
All
Language
English
Catalog #
04960
Lot #
All
Language
English
Catalog #
04900
Lot #
All
Language
English
Product Name
Collagen Solution
Catalog #
04902
Lot #
All
Language
English
Document Type
Technical Manual
Catalog #
04960
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
04960
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
04960
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
04960
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
04900
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
Collagen Solution
Catalog #
04902
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Resources and Publications

Frequently Asked Questions

Why is the MegaCult™-C formulation serum free?

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.

Why use semi-solid media?

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.

Publications (30)

The EMT regulator Zeb2/Sip1 is essential for murine embryonic hematopoietic stem/progenitor cell differentiation and mobilization. Goossens S et al. Blood 2011 MAY

Abstract

Zeb2 (Sip1/Zfhx1b) is a member of the zinc-finger E-box-binding (ZEB) family of transcriptional repressors previously demonstrated to regulate epithelial-to-mesenchymal transition (EMT) processes during embryogenesis and tumor progression. We found high Zeb2 mRNA expression levels in HSCs and hematopoietic progenitor cells (HPCs), and examined Zeb2 function in hematopoiesis through a conditional deletion approach using the Tie2-Cre and Vav-iCre recombination mouse lines. Detailed cellular analysis demonstrated that Zeb2 is dispensable for hematopoietic cluster and HSC formation in the aorta-gonadomesonephros region of the embryo, but is essential for normal HSC/HPC differentiation. In addition, Zeb2-deficient HSCs/HPCs fail to properly colonize the fetal liver and/or bone marrow and show enhanced adhesive properties associated with increased β1 integrin and Cxcr4 expression. Moreover, deletion of Zeb2 resulted in embryonic (Tie2-Cre) and perinatal (Vav-icre) lethality due to severe cephalic hemorrhaging and decreased levels of angiopoietin-1 and, subsequently, improper pericyte coverage of the cephalic vasculature. These results reveal essential roles for Zeb2 in embryonic hematopoiesis and are suggestive of a role for Zeb2 in hematopoietic-related pathologies in the adult.
Expression level and differential JAK2-V617F-binding of the adaptor protein Lnk regulates JAK2-mediated signals in myeloproliferative neoplasms. Baran-Marszak F et al. Blood 2010 DEC

Abstract

Activating mutations in signaling molecules, such as JAK2-V617F, have been associated with myeloproliferative neoplasms (MPNs). Mice lacking the inhibitory adaptor protein Lnk display deregulation of thrombopoietin/thrombopoietin receptor signaling pathways and exhibit similar myeloproliferative characteristics to those found in MPN patients, suggesting a role for Lnk in the molecular pathogenesis of these diseases. Here, we showed that LNK levels are up-regulated and correlate with an increase in the JAK2-V617F mutant allele burden in MPN patients. Using megakaryocytic cells, we demonstrated that Lnk expression is regulated by the TPO-signaling pathway, thus indicating an important negative control loop in these cells. Analysis of platelets derived from MPN patients and megakaryocytic cell lines showed that Lnk can interact with JAK2-WT and V617F through its SH2 domain, but also through an unrevealed JAK2-binding site within its N-terminal region. In addition, the presence of the V617F mutation causes a tighter association with Lnk. Finally, we found that the expression level of the Lnk protein can modulate JAK2-V617F-dependent cell proliferation and that its different domains contribute to the inhibition of multilineage and megakaryocytic progenitor cell growth in vitro. Together, our results indicate that changes in Lnk expression and JAK2-V617F-binding regulate JAK2-mediated signals in MPNs.
Role for MKL1 in megakaryocytic maturation. Cheng E-C et al. Blood 2009 MAR

Abstract

Megakaryoblastic leukemia 1 (MKL1), identified as part of the t(1;22) translocation specific to acute megakaryoblastic leukemia, is highly expressed in differentiated muscle cells and promotes muscle differentiation by activating serum response factor (SRF). Here we show that Mkl1 expression is up-regulated during murine megakaryocytic differentiation and that enforced overexpression of MKL1 enhances megakaryocytic differentiation. When the human erythroleukemia (HEL) cell line is induced to differentiate with 12-O-tetradecanoylphorbol 13-acetate, overexpression of MKL1 results in an increased number of megakaryocytes with a concurrent increase in ploidy. MKL1 overexpression also promotes megakaryocytic differentiation of primary human CD34(+) cells cultured in the presence of thrombopoietin. The effect of MKL1 is abrogated when SRF is knocked down, suggesting that MKL1 acts through SRF. Consistent with these findings in human cells, knockout of Mkl1 in mice leads to reduced platelet counts in peripheral blood, and reduced ploidy in bone marrow megakaryocytes. In conclusion, MKL1 promotes physiologic maturation of human and murine megakaryocytes.

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