Process for isolating vascular endothelial cells from embryoid bodies differentiated from embryonc stem cells
The present invention provides a process for isolating vascular endothelial cells from embryoid bodies differentiated from embryonic stem cells, which comprises: (a) treating embryoid bodies differentiated from embryonic stem cells with 0.005-0.015% trypsin and 0.05-0.15 mM ethylenediaminetetraacetate (EDTA) to obtain vascular endothelial cell clusters; and (b) treating the vascular endothelial cell clusters with 0.1-0.5% trypsin and 0.5-2 mM EDTA so as to separate the vascular endothelial cell clusters into single cells.
1. A process for isolating vascular endothelial cells from embryoid bodies differentiated from embryonic stem cells, which comprises:
(a) treating embryoid bodies differentiated from embryonic stem cells with 0.005-0.015% trypsin and 0.05-0.15 mM ethylenediaminetetraacetate (EDTA) to obtain vascular endothelial cell clusters; and then
(b) treating the vascular endothelial cell clusters with 0.1-0.5% trypsin and 0.5-2 mM EDTA so as to separate the vascular endothelial cell clusters into single cells.
2. The process of claim 1 , wherein the embryonic stem cells are human embryonic stem cells.
3. The process of claim 1 , wherein in step (a), the concentration of trypsin is 0.01% and the concentration of EDTA is 0.1 mM.
4. The process of claim 1 , wherein in step (b), the concentration of trypsin is 0.25% and the concentration of EDTA is 1 mM.
5. The process of claim 1 , wherein in steps (a) and (b), the treating with trypsin and EDTA is performed for 3 to 10 minutes, respectively.
6. The process of claim 2 , wherein in step (a), the concentration of trypsin is 0.01% and the concentration of EDTA is 0.1 mM.
7. The process of claim 2 , wherein in step (b), the concentration of trypsin is 0.25% and the concentration of EDTA is 1 mM.
8. A process for isolating vascular endothelial cells from embryoid bodies differentiated from embryonic stem cells, which comprises:
(a) treating embryoid bodies differentiated from embryonic stem cells with 0.005-0.015% trypsin and 0.05-0.15 mM ethylenediaminetetraacetate (EDTA) to obtain vascular endothelial cell clusters; and
(b) treating the vascular endothelial cell clusters with 0.1-0.5% trypsin and 0.5-2 mM EDTA so as to separate the vascular endothelial cell clusters into single cells;
wherein the embryonic stem cells are human embryonic stem cells; and
wherein in steps (a) and (b), trypsin and EDTA are treated for 3 to 10 minutes.