Label-free identification of stem cell-differentiated cells

Abstract

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 system comprising (a) a laser light source to produce an incident light, (b) a sensor configured to collect or detect second-harmonic light generated from the incident light, and (c) a plurality of cells comprising at least an undifferentiated stem cell and a differentiated cell comprising a noncentrosymmetric structure. 
     
     
         2 . The system of  claim 1 , wherein the noncentrosymmetric structure is one or more of myosin, collagen, microtubule, axon or dendrite. 
     
     
         3 . The system of  claim 1 , wherein the differentiated cell was differentiated in vitro from a stem cell for a duration of less than 180 days. 
     
     
         4 . The system of  claim 3 , wherein the duration of the differentiation was less than 30 days. 
     
     
         5 . The system of  claim 1 , wherein the differentiated cell is selected from an immature cardiomyocyte, an immature skeleton muscle cell, an immature smooth muscle cell or an immature neuron. 
     
     
         6 . The system of  claim 1 , wherein the differentiated cell is an immature cardiomyocyte. 
     
     
         7 . The system of  claim 3 , wherein the stem cell that produces the a stem-cell derived differentiated cell 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. 
     
     
         8 . The system of  claim 1 , wherein the incident light has a wavelength from about 700 nm to about 1500 nm. 
     
     
         9 . The system of  claim 1 , wherein the incident light comprises pulses having a width of about 70 fs to about 10 ps. 
     
     
         10 . The system of  claim 1 , wherein the incident light has a repetition rate from about 1 KHz to about 100 MHz. 
     
     
         11 . The system of  claim 1 , wherein the cells do not comprise an exogenous label. 
     
     
         12 . The system of  claim 11 , wherein the exogenous label is a fluorescent label or a radioactive label. 
     
     
         13 . The system of  claim 1 , wherein the cells are suspended in a medium. 
     
     
         14 . The system of  claim 13 , wherein the medium comprises collagenase. 
     
     
         15 . The system of  claim 13 , wherein the medium does not contain trypsin. 
     
     
         16 . The system of  claim 1 , wherein the stem cell is a mammalian stem cell. 
     
     
         17 . The system of  claim 16 , wherein the mammalian stem cell is of the group of a murine stem cell or a human stem cell.