CD309 (Flk-1) MicroBead Kit, mouse

Name: CD309 (Flk-1) MicroBead Kit, mouse
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CD309 (Flk-1) MicroBead Kit, mouse

The CD309 (Flk-1) MicroBead Kit has been developed for the isolation of CD309 (Flk-1)

⁺

cells from differentiated murine ES or iPS cell cultures. Isolation of CD309

⁺

cells from heterogeneous differentiation cultures can improve differentiation into specific hematopoietic or cardiovascular lineages, e.g. endothelial cells.

Product data and images for CD309 (Flk-1) MicroBead Kit, mouse

Figures

Figure 1

Mouse ES cells (HM1) were differentiated into embryoid bodies (EBs). At day 5 EBs were dissociated with the Embryoid Body Dissociation Kit (# 130-096-348) and CD309 expressing cells were isolated using the CD309 (Flk-1) MicroBead Kit, an LS Column, and a MidiMACS™ Separator. After fluorescent labeling with Labeling Check Reagent‑APC (# 130-095-237) cells were analyzed by flow cytometry using the MACSQuant

^®

Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Unseparated cells

Isolated CD309

⁺

cells

Figure 1

Mouse ES cells (HM1) were differentiated into embryoid bodies (EBs). At day 5 EBs were dissociated with the Embryoid Body Dissociation Kit (# 130-096-348) and CD309 expressing cells were isolated using the CD309 (Flk-1) MicroBead Kit, an LS Column, and a MidiMACS™ Separator. After fluorescent labeling with Labeling Check Reagent‑APC (# 130-095-237) cells were analyzed by flow cytometry using the MACSQuant

^®

Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Figure 1

Mouse ES cells (HM1) were differentiated into embryoid bodies (EBs). At day 5 EBs were dissociated with the Embryoid Body Dissociation Kit (# 130-096-348) and CD309 expressing cells were isolated using the CD309 (Flk-1) MicroBead Kit, an LS Column, and a MidiMACS™ Separator. After fluorescent labeling with Labeling Check Reagent‑APC (# 130-095-237) cells were analyzed by flow cytometry using the MACSQuant

^®

Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Figure 2

Improved endothelial differentiation of isolated CD309 (Flk-1)⁺ cells:

Unseparated or CD309-enriched cardiovascular progenitor cells (derived as in fig. 1) were subjected to endothelial differentiation in the presence of 50 ng/mL VEGF. Differentiation efficiency was evaluated by immunofluorescence using CD31-FITC (green) and DAPI counterstain (blue).

Unseparated cells

Isolated CD309

⁺

cells

Figure 2

Improved endothelial differentiation of isolated CD309 (Flk-1)⁺ cells:

Unseparated or CD309-enriched cardiovascular progenitor cells (derived as in fig. 1) were subjected to endothelial differentiation in the presence of 50 ng/mL VEGF. Differentiation efficiency was evaluated by immunofluorescence using CD31-FITC (green) and DAPI counterstain (blue).

Figure 2

Improved endothelial differentiation of isolated CD309 (Flk-1)⁺ cells:

Unseparated or CD309-enriched cardiovascular progenitor cells (derived as in fig. 1) were subjected to endothelial differentiation in the presence of 50 ng/mL VEGF. Differentiation efficiency was evaluated by immunofluorescence using CD31-FITC (green) and DAPI counterstain (blue).

Specifications for CD309 (Flk-1) MicroBead Kit, mouse

Overview

The CD309 (Flk-1) MicroBead Kit has been developed for the isolation of CD309 (Flk-1)

⁺

cells from differentiated murine ES or iPS cell cultures. Isolation of CD309

⁺

cells from heterogeneous differentiation cultures can improve differentiation into specific hematopoietic or cardiovascular lineages, e.g. endothelial cells.

Detailed product information

Background information

CD309 is a class 3 receptor tyrosine kinase, also known as Flk-1 (fetal liver kinase 1), vascular endothelial growth factor receptor 2 (VEGFR2), or kinase insert domain receptor (KDR). CD309 is one of the earliest differentiation markers for endothelial and hematopoietic cells.

^1,2

Using mouse embryonic stem cells it has been shown that CD309 is a marker for multipotent cardiovascular progenitor cells that give rise to cardiomyocyte, endothelial, and vascular smooth muscle cell lineages.

^3,4

Note

: For optimal flow cytometric analysis of isolated cells after separation we recommend Labeling Check Reagent.

Columns

LS or autoMACS

^®

Columns.

References for CD309 (Flk-1) MicroBead Kit, mouse

Publications

Shalaby, F. et al. (1995) Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature 376: 62-66
Eichmann, A. et al. (1997) Ligand-dependent development of the endothelial and hemopoietic lineages from embryonic Proc. Natl. Acad. Sci. U.S.A. 94: 5141-5146
Kattmann et al. (2006) Dev. Cell 5: 723-732
Yamashita, J. et al. (2000) Flk1-positive cells derived from embryonic stem cells serve as vascular progenitors. Nature 408(6808): 92-96

Data and images