Method of screening endothelial cells for angiogenic capability
View Patent ↗Provided is a method of screening a primary endothelial cell population for angiogenesis capability comprising: (a) measuring the percentage of cells that are positive for VEGF R2 and CD34, the level of VEGF R2, or measuring the VEGF R2 to VEGF R1 ratio in the population; and (b) selecting those populations where the measured percentage or the measured ratio is over a threshold value.
1. A method of selecting for higher angiogenesis capability among endothelial cells in an endothelial cell population, said method comprising:
a) staining the endothelial cell population with anti-VEGF R2 and anti-VEGF R1;
b) measuring the VEGF R2 to VEGF R1 ratio in the stained population; and
c) selecting endothelial cells having a higher VEGF R2 to VEGF R1 ratio wherein the higher the VEGF R2 to VEGF R1 ratio, the higher the capability of the endothelial cells to form tubes and the higher the angiogenic capability of the endothelial cells.
2. The method of claim 1 , wherein the step of measuring the VEGF R2 to VEGF R1 ratio in the stained population comprises fluorescence activated cell sorting analysis.
3. The method of claim 1 , wherein the step of measuring the VEGF R2 to VEGF R1 ratio in the stained population comprises immunofluorescent imaging and data processing.
4. The method of claim 1 , wherein the step of measuring the VEGF R2 to VEGF R1 ratio in the stained population comprises a cell-based Enzyme-Linked Immunosorbent Assay.
5. The method of claim 1 , further comprising labeling the endothelial cell population with one or more secondary antibodies.
6. The method of claim 5 , wherein the one or more secondary antibodies are fluorescein or rhodamine conjugated.
7. The method of claim 1 , wherein the endothelial cell population comprises human dermal microvascular endothelial cells.
8. The method of claim 1 , wherein the endothelial cell population comprises human pulmonary microvascular endothelial cells.
9. The method of claim 1 , wherein the endothelial cell population comprises human umbilical vein endothelial cells.