Document Detail

Tracking stem cell differentiation in the setting of automated optogenetic stimulation.
MedLine Citation:
PMID:  21280159     Owner:  NLM     Status:  MEDLINE    
Membrane depolarization has been shown to play an important role in the neural differentiation of stem cells and in the survival and function of mature neurons. Here, we introduce a microbial opsin into ESCs and develop optogenetic technology for stem cell engineering applications, with an automated system for noninvasive modulation of ESC differentiation employing fast optogenetic control of ion flux. Mouse ESCs were stably transduced with channelrhodopsin-2 (ChR2)-yellow fluorescent protein and purified by fluorescence activated cell sorting (FACS). Illumination of resulting ChR2-ESCs with pulses of blue light triggered inward currents. These labeled ESCs retained the capability to differentiate into functional mature neurons, assessed by the presence of voltage-gated sodium currents, action potentials, fast excitatory synaptic transmission, and expression of mature neuronal proteins and neuronal morphology. We designed and tested an apparatus for optically stimulating ChR2-ESCs during chronic neuronal differentiation, with high-speed optical switching on a custom robotic stage with environmental chamber for automated stimulation and imaging over days, with tracking for increased expression of neural and neuronal markers. These data point to potential uses of ChR2 technology for chronic and temporally precise noninvasive optical control of ESCs both in vitro and in vivo, ranging from noninvasive control of stem cell differentiation to causal assessment of the specific contribution of transplanted cells to tissue and network function.
Albrecht Stroh; Hsing-Chen Tsai; Li-Ping Wang; Feng Zhang; Jenny Kressel; Alexander Aravanis; Nandhini Santhanam; Karl Deisseroth; Arthur Konnerth; M Bret Schneider
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Stem cells (Dayton, Ohio)     Volume:  29     ISSN:  1549-4918     ISO Abbreviation:  Stem Cells     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2011-01-31     Completed Date:  2011-05-02     Revised Date:  2014-10-03    
Medline Journal Info:
Nlm Unique ID:  9304532     Medline TA:  Stem Cells     Country:  United States    
Other Details:
Languages:  eng     Pagination:  78-88     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 AlphaMed Press.
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MeSH Terms
Action Potentials
Cell Tracking / instrumentation*,  methods*
Central Nervous System Diseases / surgery
Embryonic Stem Cells / cytology*,  metabolism
Gene Expression Profiling
Image Processing, Computer-Assisted
Induced Pluripotent Stem Cells / cytology*,  metabolism,  transplantation
Microscopy, Confocal
Neurons / cytology*,  metabolism,  physiology
Rats, Wistar
Recombinant Fusion Proteins / genetics,  metabolism
Rhodopsin / genetics,  metabolism
Stereotaxic Techniques
Grant Support
Reg. No./Substance:
0/Recombinant Fusion Proteins; 0/channelrhodopsin 2, mouse; 9009-81-8/Rhodopsin

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