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MegaCult™-C Collagen and Medium with Cytokines

Collagen and medium with cytokines for human CFU-Mk assays
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产品号 #04961_C

Collagen and medium with cytokines for human CFU-Mk assays

产品优势


  • Serum-free formulation

产品组分包括

  • Collagen Solution, 35 mL
  • MegaCult™-C Medium with Cytokines, 24 x 2 mL (Catalog #04901)
main product photo
MegaCult™-C Collagen and Medium with Cytokines
概述
产品说明书及文档
应用领域
相关材料与文献

概述

MegaCult™-C Collagen and Medium with Cytokines kit includes includes medium and collagen solution necessary for performing colony-forming unit (CFU) assays of human megakaryocyte progenitor cells (CFU-Mk).

• MegaCult™-C Medium with Cytokines is optimized for the growth of CFU-Mk in human bone marrow, mobilized peripheral blood, and cord blood samples. It is suitable for use with CD34 + enriched cells, mononuclear cells, and cells isolated by other purification methods. The medium contains recombinant human (rh) IL-3, IL-6, and thrombopoietin.
• Collagen Solution for preparation of collagen gels and for coating cell culture surfaces.
Individual kit components may also be purchased separately, for your convenience.

For more information on protocols for human CFU-Mk assays using MegaCult™-C, please explore the Technical Manual.

Subtype
Semi-Solid Media, Specialized Media
 
Cell Type
Hematopoietic Stem and Progenitor Cells
 
Species
Human
 
Application
Cell Culture, Colony Assay, Functional Assay
 
Brand
MegaCult
 
Area of Interest
Stem Cell Biology
 
Formulation Category
Serum-Free
 

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
Document Type
Product Information Sheet 1
Product Name
MegaCult™-C Collagen and Medium with Cytokines
Catalog #
04961
Lot #
All
Language
English
Document Type
Product Information Sheet 2
Product Name
MegaCult™-C Collagen and Medium with Cytokines
Catalog #
04961
Lot #
All
Language
English
Document Type
Technical Manual
Product Name
MegaCult™-C Collagen and Medium with Cytokines
Catalog #
04961
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
MegaCult™-C Collagen and Medium with Cytokines
Catalog #
04961
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
MegaCult™-C Collagen and Medium with Cytokines
Catalog #
04961
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.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (1)

Hematopoietic Stem and Progenitor Cells - Products for Your Research
Brochure
Hematopoietic Stem and Progenitor Cells - Products for Your Research

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 (23)

Mef2C is a lineage-restricted target of Scl/Tal1 and regulates megakaryopoiesis and B-cell homeostasis. Gekas C et al. Blood 2009 APR

Abstract

The basic helix-loop-helix transcription factor stem cell leukemia gene (Scl) is a master regulator for hematopoiesis essential for hematopoietic specification and proper differentiation of the erythroid and megakaryocyte lineages. However, the critical downstream targets of Scl remain undefined. Here, we identified a novel Scl target gene, transcription factor myocyte enhancer factor 2 C (Mef2C) from Scl(fl/fl) fetal liver progenitor cell lines. Analysis of Mef2C(-/-) embryos showed that Mef2C, in contrast to Scl, is not essential for specification into primitive or definitive hematopoietic lineages. However, adult VavCre(+)Mef2C(fl/fl) mice exhibited platelet defects similar to those observed in Scl-deficient mice. The platelet counts were reduced, whereas platelet size was increased and the platelet shape and granularity were altered. Furthermore, megakaryopoiesis was severely impaired in vitro. Chromatin immunoprecipitation microarray hybridization analysis revealed that Mef2C is directly regulated by Scl in megakaryocytic cells, but not in erythroid cells. In addition, an Scl-independent requirement for Mef2C in B-lymphoid homeostasis was observed in Mef2C-deficient mice, characterized as severe age-dependent reduction of specific B-cell progenitor populations reminiscent of premature aging. In summary, this work identifies Mef2C as an integral member of hematopoietic transcription factors with distinct upstream regulatory mechanisms and functional requirements in megakaryocyte and B-lymphoid lineages.
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Transgenic expression of JAK2V617F causes myeloproliferative disorders in mice. Xing S et al. Blood 2008 MAY

Abstract

The JAK2(V617F) mutation was found in most patients with myeloproliferative disorders (MPDs), including polycythemia vera, essential thrombocythemia, and primary myelofibrosis. We have generated transgenic mice expressing the mutated enzyme in the hematopoietic system driven by a vav gene promoter. The mice are viable and fertile. One line of the transgenic mice, which expressed a lower level of JAK2(V617F), showed moderate elevations of blood cell counts, whereas another line with a higher level of JAK2(V617F) expression displayed marked increases in blood counts and developed phenotypes that closely resembled human essential thrombocythemia and polycythemia vera. The latter line of mice also developed primary myelofibrosis-like symptoms as they aged. The transgenic mice showed erythroid, megakaryocytic, and granulocytic hyperplasia in the bone marrow and spleen, displayed splenomegaly, and had reduced levels of plasma erythropoietin and thrombopoietin. They possessed an increased number of hematopoietic progenitor cells in peripheral blood, spleen, and bone marrow, and these cells formed autonomous colonies in the absence of growth factors and cytokines. The data show that JAK2(V617F) can cause MPDs in mice. Our study thus provides a mouse model to study the pathologic role of JAK2(V617F) and to develop treatment for MPDs.
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Increased production of megakaryocytes near purity from cord blood CD34+ cells using a short two-phase culture system. Boyer L et al. Journal of immunological methods 2008 MAR

Abstract

Expansion of hematopoietic progenitor cells (HPC) ex vivo remains an important focus in fundamental and clinical research. The aim of this study was to determine whether the implementation of such expansion phase in a two-phase culture strategy prior to the induction of megakaryocyte (Mk) differentiation would increase the yield of Mks produced in cultures. Toward this end, we first characterized the functional properties of five cytokine cocktails to be tested in the expansion phase on the growth and differentiation kinetics of CD34+-enriched cells, and on their capacity to expand clonogenic progenitors in cultures. Three of these cocktails were chosen based on their reported ability to induce HPC expansion ex vivo, while the other two represented new cytokine combinations. These analyses revealed that none of the cocktails tested could prevent the differentiation of CD34+ cells and the rapid expansion of lineage-positive cells. Hence, we sought to determine the optimum length of time for the expansion phase that would lead to the best final Mk yields. Despite greater expansion of CD34+ cells and overall cell growth with a longer expansion phase, the optimal length for the expansion phase that provided greater Mk yield at near maximal purity was found to be 5 days. Under such settings, two functionally divergent cocktails were found to significantly increase the final yield of Mks. Surprisingly, these cocktails were either deprived of thrombopoietin or of stem cell factor, two cytokines known to favor megakaryopoiesis and HPC expansion, respectively. Based on these results, a short resource-efficient two-phase culture protocol for the production of Mks near purity (textgreater95%) from human CD34+ CB cells has been established.
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