Label-free identification of stem cell-differentiated cells
View Patent ↗Methods are disclosed for isolation of a stem-cell derived differentiated cell, which method entails illuminating an incident light onto a plurality of stem-cell derived cells that comprise at least an undifferentiated cell and a differentiated cell possessing a noncentrosymmetric structure, wherein the differentiated cell generates second-harmonic light from the incident light; and isolating the differentiated cell identified by the second-harmonic light. Devices for carrying out the methods are also provided.
1. A method for sorting and isolating a stem-cell derived, stage-specific immature cardiomyocyte, comprising:
(a) illuminating an incident light onto a plurality of stem-cell derived cells that comprise at least an undifferentiated cell and an immature cardiomyocyte possessing an intracellular noncentrosymmetric structure, wherein the immature cardiomyocyte generates second-harmonic light from the incident light;
(b) identifying the stage of differentiation in the immature cardiomyocyte by detecting the intensity of the second-harmonic light generated by the immature cardiomyocyte; and
(c) sorting and isolating the stage-specific immature cardiomyocyte identified in step (b), wherein the noncentrosymmetric structure is myosin.
2. The method of claim 1 , wherein the immature cardiomyocyte is differentiated, in vitro, from a stem-cell wherein the duration of the differentiation is less than 180 days.
3. The system of claim 2 , wherein the duration of the differentiation is less than 30 days.
4. The method of claim 2 , wherein the stem-cell that produces the a stem-cell derived immature cardiomyocyte is of the group: an embryonic stem cell, an induced pluripotent stem cell, a multipotent stem cell, an oligopotent stem cell, or a unipotent stem cell.
5. The method of claim 1 , wherein the incident light has a wavelength from about 700 nm to about 1500 nm.
6. The method of claim 1 , wherein the incident light comprises pulses having a width of about 70 fs to about 10 ps.
7. The method of claim 1 , wherein the incident light has a repetition rate from about 1 KHz to about 100 MHz.
8. The method of claim 1 , wherein the cell is isolated in the absence of exogenous label or the cell is isolated in the absence of genetic modification of the cells.
9. The method of claim 8 , wherein the exogenous label is a fluorescent label or a radioactive label.
10. The method of claim 1 , further comprising suspending the cells in a medium prior to illuminating the incident light onto the cells.
11. The method of claim 10 , wherein the medium comprises collagenase.
12. The method of claim 10 , wherein the medium does not contain trypsin.
13. The method of claim 1 , wherein the stem cell is a mammalian stem cell.
14. The method or population of claim 13 , wherein the stem cell is of the group of a murine stem cell or a human stem cell.
15. The method of claim 1 , wherein the stage of differentiation is identified by detecting low intensity second harmonic light.
16. The method of claim 1 , wherein the stage of differentiation is identified by detecting high intensity second harmonic light.