Integrative genomic and functional analyses reveal neuronal subtype differentiation bias in human embryonic stem cell lines

doi:10.1073/pnas.0706199104

Integrative genomic and functional analyses reveal neuronal subtype differentiation bias in human embryonic stem cell lines

*Mental Retardation Research Center and
^†Departments of Psychiatry and Biobehavioral Sciences and Molecular and Medical Pharmacology, Neuropsychiatric Institute, David Geffen School of Medicine at University of California, Los Angeles, Neuroscience Research Building Room 351, 635 Charles E. Young Drive South, Los Angeles, CA 90095;
^‡Departments of Neuroscience and Molecular Genetics and
^§Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, 6000 Harry Hines Boulevard NA4.118, Dallas, TX 75390; and
^¶Applied Biosystems, Inc., Foster City, CA 94404

Contributed by Thomas C. Südhof, July 2, 2007 (received for review June 24, 2007)

Abstract

The self-renewal and differentiation potential of human embryonic stem cells (hESCs) suggests that hESCs could be used for regenerative medicine, especially for restoring neuronal functions in brain diseases. However, the functional properties of neurons derived from hESC are largely unknown. Moreover, because hESCs were derived under diverse conditions, the possibility arises that neurons derived from different hESC lines exhibit distinct properties, but this possibility remains unexplored. To address these issues, we developed a protocol that allows stepwise generation from hESCs of cultures composed of ≈70–80% human neurons that exhibit spontaneous synaptic network activity. Comparison of neurons derived from the well characterized HSF1 and HSF6 hESC lines revealed that HSF1- but not HSF6-derived neurons exhibit forebrain properties. Accordingly, HSF1-derived neurons initially form primarily GABAergic synaptic networks, whereas HSF6-derived neurons initially form glutamatergic networks. microRNA profiling revealed significant expression differences between the two hESC lines, suggesting that microRNAs may influence their distinct differentiation properties. These observations indicate that although both HSF1 and HSF6 hESCs differentiate into functional neurons, the two hESC lines exhibit distinct differentiation potentials, suggesting that they are preprogrammed. Information on hESC line-specific differentiation biases is crucial for neural stem cell therapy and establishment of novel disease models using hESCs.

Footnotes

^‖To whom correspondence may be addressed. E-mail: thomas.sudhof{at}utsouthwestern.edu or ysun{at}mednet.ucla.edu
Author contributions: J.X., Z.P.P., and W.G. contributed equally to this work; H.W., J.X., Z.P.P., T.C.S., and Y.E.S. designed research; H.W., J.X., Z.P.P., W.G., K.J.K., B.B., and Y.E.S. performed research; C.C. contributed new reagents/analytic tools; H.W., J.X., Z.P.P., W.G., K.J.K., T.C.S., and Y.E.S. analyzed data; and H.W., J.X., Z.P.P., T.C.S., and Y.E.S. wrote the paper.
The authors declare no conflict of interest.
This article contains supporting information online at www.pnas.org/cgi/content/full/0706199104/DC1.
Abbreviations:

EPSC,

excitatory postsynaptic current;

hNPCs,

human neural stem/progenitor cell;

hESC,

human embryonic stem cell;

IPSC,

inhibitory postsynaptic current.