Adhesive signature-based methods for the isolation of stem cells and cells derived therefrom
The present invention provides for methods of isolating a stem cell or cell derived therefrom from a mixture of cells, for example, a mixture of adherent cells in culture. Cell isolation is achieved by the application of selective detachment forces.
1. A method of isolating a human induced pluripotent stem cell (hiPSC) and/or a human embryonic stem cell (hESC) from fibroblasts adhered to a substrate:
a) growing a mixture of cells comprising fibroblasts and an hiPSC and/or an hESC on a substrate in culture, such that the cells in the mixture of cells are adhered to the substrate; and
b) subjecting the mixture of adhered cells comp rising fibroblasts and an hiPSC and/or an hESC to a detachment force that provides a wall shear stress in the range from about 70 to about 160 dynes/cm 2 , wherein the detachment force selectively detaches the hiPSC and/or hESC from the mixture of adhered cells, thereby isolating the hiPSC and/or hESC from the mixture of cells adhered to the substrate.
2. The method of claim 1 , wherein the detachment force that is sufficient to selectively detach the hiPSC and/or hESC provides a wall shear stress in the range from about 85 to about 125 dynes/cm 2 .
3. The method of claim 1 , wherein the method further comprises culturing the isolated hiPSC and/or hESC.
4. The method of claim 1 , wherein the method further comprises evaluating the isolated hiPSC and/or hESC by flow cytometry, biochemical analysis and/or gene expression analysis.
5. The method of claim 1 , wherein the method does not comprise attaching a detectable label and/or affinity reagent to the mixture of animal cells.
6. The method of claim 1 , wherein the detachment force is applied by hydrodynamic force, centrifugal force and/or magnetic force.
7. The method of claim 1 , wherein the method is carried out in a microfluidic device.
8. The method of claim 1 , wherein the mixture of cultured cells is subjected to the detachment force for 1 to 60 minutes.
9. The method of claim 1 , wherein the mixture of cultured cells is subjected to the detachment force for 2 to 20 minutes.