At the time of submission of the orginal ALS iPS Go Grant, there were no well-validated protocols for generating astroglia from human ES or IPS cell lines, although approaches from rodent cells existed. Fortunately, we were able to develop a reliable protocol in both the Henderson lab and the Rothstein labs (Figure 5A). We now have a protocol that works identically in both labs?with very similar efficiency at differentiation between labs. We have also been able to detect evidence of true astroglial maturation in a number of lines that have advanced sufficiently and these express appropriate astroglial specific proteins and have functional glutamate transport (Figure 5B-D).

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Resource-Related Research Projects--Cooperative Agreements (U24)
Project #
1U24NS078736-01
Application #
8293969
Study Section
Special Emphasis Panel (ZNS1-SRB-S (53))
Project Start
2012-07-01
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
1
Fiscal Year
2012
Total Cost
$849,613
Indirect Cost
$218,153
Name
Johns Hopkins University
Department
Type
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Zhang, Ke; Daigle, J Gavin; Cunningham, Kathleen M et al. (2018) Stress Granule Assembly Disrupts Nucleocytoplasmic Transport. Cell 173:958-971.e17
Zhang, Ping-Wu; Haidet-Phillips, Amanda M; Pham, Jacqueline T et al. (2016) Generation of GFAP::GFP astrocyte reporter lines from human adult fibroblast-derived iPS cells using zinc-finger nuclease technology. Glia 64:63-75
Zhang, Ke; Donnelly, Christopher J; Haeusler, Aaron R et al. (2015) The C9orf72 repeat expansion disrupts nucleocytoplasmic transport. Nature 525:56-61
Li, Ying; Balasubramanian, Umamahesw; Cohen, Devon et al. (2015) A comprehensive library of familial human amyotrophic lateral sclerosis induced pluripotent stem cells. PLoS One 10:e0118266
Wainger, Brian J; Kiskinis, Evangelos; Mellin, Cassidy et al. (2014) Intrinsic membrane hyperexcitability of amyotrophic lateral sclerosis patient-derived motor neurons. Cell Rep 7:1-11
Kiskinis, Evangelos; Sandoe, Jackson; Williams, Luis A et al. (2014) Pathways disrupted in human ALS motor neurons identified through genetic correction of mutant SOD1. Cell Stem Cell 14:781-95