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.

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
View details

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.
View details

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
View details

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).
View details

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

  1. Shalaby, F. et al. (1995) Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature 376: 62-66
  2. 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
  3. Kattmann et al. (2006) Dev. Cell 5: 723-732
  4. Yamashita, J. et al. (2000) Flk1-positive cells derived from embryonic stem cells serve as vascular progenitors. Nature 408(6808): 92-96

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