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products
EndoCult® Liquid Medium Kit
EndoCult® Colony Atlas
background
Blood vessel development is a regulated process involving the proliferation and migration of endothelial cells from adjacent pre-existing blood vessels (angiogenesis), or the differentiation of endothelial progenitor cells (EPCs) from mesodermal precursors (vasculogenesis).1,2 EPCs or angioblasts were originally thought to be present only during embryonic development, however, evidence accumulated in the past several years suggest that they can persist in adult life.1,2 This has generated interest in the use of EPCs for neovascularization of ischemic or injured tissue and for the clinical assessment of risk factors for various diseases.3-5 More recently, populations of cells that support or augment angiogenesis and/or vasculogenesis, without actualy differentiating into cells that form part of the vascular network, have also been identified.
5 Day CFU-Hill Colony Assay
Hill et al.3 developed a cell culture assay to assess the correlation between the frequency of a specific population of putative circulating EPCs and multiple clinical factors. This in vitro assay has been standardized by STEMCELL Technologies and is referred to as the 5 Day CFU-Hill Colony Assay. The EndoCult® Liquid Medium Kit has been developed specifically to support the growth and quantification of CFU-Hill colonies. In this protocol, mononuclear cells are isolated by Ficoll™ density gradient centrifugation and plated on fibronectin-coated 6-well plates in EndoCult® Medium. After 48 hours, non-adherent cells are harvested, enumerated and replated in replicate fibronectin-coated 24-well plates. CFU-Hill colonies are evaluated and quantified three days later. A CFU-Hill colony is defined as a central core of "round" cells surrounded by radiating thin flat cells. Hill et al.3, using the assay found that in healthy individuals, the number of colonies negatively correlated with Framingham cardiovascular risk score and positively correlated with vascular function, as measured by flow-mediate brachial artery reactivity.3
applications of EndoCult® and the CFU-Hill Colony Assay:
» The standardized culture and quantification of CFU-Hill colonies, which serve as a biological marker for vascular function and cardiovascular risk3
» Identification of biological factors inhibiting or promoting colony formation, which may potentially also influence vascular function and cardiovascular risk
» Functional analysis for drug screening or dissection of cellular pathways
» Analysis of cells or factors involved in angiogenesis or vasculogenesis
recent reports that used the 5 Day CFU-Hill Colony Assay and/or EndoCult® in investigations concerning colony frequency and:
» coronary artery disease
» peripheral arterial disease
» diabetes
» chronic obstructive pulmonary disease
» asthma
» hypogonadism
» stroke
FOR RESEARCH USE ONLY
Also available: ES-Cult™ Endothelial products for the in vitro differentiation of mouse ES cells into endothelial cells.
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