Optically controllable fgfr stimulation using wireless controlled cellular lighting system

Abstract

The present invention relates to the field of stem cells. More specifically, the present invention provides compositions and methods for using optogenetics to sustain the pluripotency of stem cells. In one embodiment, a vector comprises a nucleotide sequencing encoding a fusion protein comprising the intracellular domain of fibroblast growth factor 1 receptor (FGFR1) and a photoactivatable domain.

1 - 33 . (canceled) 
     
     
         34 . A method for maintaining cells, the method comprising:
 (a) providing or obtaining, in a culture medium, cells genetically engineered to be maintained by exposure to light; and   (b) illuminating the cells with light at a sufficient light intensity and time such that the cells are maintained, thereby maintaining the cells,   
       wherein the method does not require a step of supplementing the culture medium with at least one growth factor required for maintaining the cells. 
     
     
         35 . The method of  claim 34 , wherein the illuminating of (b) replaces the step of supplementing the culture medium with the at least one growth factor required for maintaining the cells. 
     
     
         36 . The method of  claim 34 , wherein the cells provided or obtained are capable of self-renewal. 
     
     
         37 . The method of  claim 36 , wherein, after maintaining the cells, the cells are capable of self-renewal. 
     
     
         38 . The method of  claim 34 , wherein the cells provided or obtained have the potential to differentiate or to further differentiate into different cell types. 
     
     
         39 . The method of  claim 38 , wherein, after maintaining the cells, the cells have the potential to differentiate or to further differentiate into different cell types. 
     
     
         40 . The method of  claim 34 , wherein the cells provided or obtained are stem cells. 
     
     
         41 . The method of  claim 40 , wherein the stem cells are pluripotent stem cells. 
     
     
         42 . The method of  claim 40 , wherein the stem cells are embryonic stem cells. 
     
     
         43 . The method of  claim 40 , wherein the stem cells are induced pluripotent stem cells. 
     
     
         44 . The method of  claim 40 , wherein the maintaining comprises maintaining pluripotency of the stem cells. 
     
     
         45 . The method of  claim 34 , wherein the cells provided or obtained are genetically engineered to express a fusion protein comprising a growth factor receptor protein or a portion thereof fused to a photoactivatable domain. 
     
     
         46 . The method of  claim 45 , wherein the photoactivatable domain is selected from the group consisting of: cryptochrome 2 (Cry2), light, oxygen, and voltage (LOV) domain, phytochrome B (PhyB), and UV-resistance locus 8 (UVR8). 
     
     
         47 . The method of  claim 45 , wherein the growth factor receptor protein or portion thereof is a fibroblast growth factor receptor (FGFR), a transforming growth factor receptor (TGFR), a portion thereof, or a domain thereof. 
     
     
         48 . The method of  claim 47 , wherein the growth factor receptor protein or portion thereof is FGFR or an intracellular domain of FGFR. 
     
     
         49 . The method of  claim 48 , wherein the method results in phosphorylation of ERK1/2. 
     
     
         50 . The method of  claim 48 , wherein the method does not require a step of supplementing the culture medium with exogenous fibroblast growth factor (FGF). 
     
     
         51 . The method of  claim 34 , wherein the cells are maintained for at least 1 week. 
     
     
         52 . The method of  claim 34 , wherein the cells provided or obtained are mammalian cells. 
     
     
         53 . The method of  claim 34 , wherein the cells provided or obtained are from a livestock animal. 
     
     
         54 . The method of  claim 53 , wherein the livestock animal is selected from the group consisting of: a pig, a cow, a sheep, and a goat. 
     
     
         55 . The method of  claim 34 , wherein the cells provided or obtained are from a chicken. 
     
     
         56 . The method of  claim 34 , wherein the cells provided or obtained are from a human. 
     
     
         57 . The method of  claim 34 , wherein the light is produced by an illumination source. 
     
     
         58 . The method of  claim 57 , wherein the illumination source comprises a light-emitting diode (LED). 
     
     
         59 . The method of  claim 34 , wherein the intensity is from about 0.1 μW/mm 2  to about 25 μW/mm 2 . 
     
     
         60 . The method of  claim 57 , wherein the illumination source: (i) illuminates the cells for a period of time from about 1 minute to about 120 minutes; (ii) illuminates the cells in a time interval of from about 30 minutes to about 4 hours; or (iii) both (i) and (ii). 
     
     
         61 . The method of  claim 57 , wherein the illumination source continuously illuminates the cells. 
     
     
         62 . The method of  claim 57 , wherein the illumination source illuminates the cells with a pulsed light pattern. 
     
     
         63 . The method of  claim 57 , wherein the illumination source spatiotemporally controls illumination of the cells.