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 cell of interest from a mixture of animal cells adhered to a substrate, the method comprising:
a) growing the mixture of animal cells 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 comprising the cell of interest and at least one other cell type, which is different than the cell of interest, to a detachment force that provides a wall shear stress in a range wherein the detachment force selectively detaches the cell of interest from the mixture of adhered cells, thereby isolating the cell of interest from the mixture of cells adhered to the substrate, wherein:
i) the cell of interest is a fibroblast and/or a directly differentiated or spontaneously differentiated hiPSC and the at least one other cell type is a hiPSC and/or a hESC and the detachment force provides a wall shear stress in a range of 750-850 dynes/cm 2 ;
ii) the cell of interest is a partially reprogrammed hiPSC and the at least one other cell type is a hiPSC and/or a hESC and the detachment force provides a wall shear stress in a range of 170-230 dynes/cm 2 ; and/or
iii) the cell of interest is an adult stem cell and the at least one other cell type is a hiPSC and/or a hESC and the detachment force provides a wall shear stress in a range of 480-540 dynes/cm 2 .
2. The method of claim 1 , wherein the method further comprises culturing the isolated cell of interest.
3. The method of claim 1 , wherein the method further comprises evaluating the isolated, cell of interest by flow cytometry, biochemical analysis and/or gene expression analysis.
4. 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.
5. The method of claim 1 , wherein the detachment force is applied by hydrodynamic force, centrifugal force and/or magnetic force.
6. The method of claim 1 , wherein the method is carried out in a microfluidic device.
7. The method of claim 1 , wherein the mixture of cultured cells is subjected to the detachment force for 1 to 60 minutes.
8. The method of claim 1 , wherein the mixture of cultured cells is subjected to the detachment force for 2 to 20 minutes.