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Translational and Clinical Research
Article first published online: 20 AUG 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 9, pages 2054–2062, September 2012
How to Cite
Jeon, I., Lee, N., Li, J.-Y., Park, I.-H., Park, K. S., Moon, J., Shim, S. H., Choi, C., Chang, D.-J., Kwon, J., Oh, S.-H., Shin, D. A., Kim, H. S., Do, J. T., Lee, D. R., Kim, M., Kang, K.-S., Daley, G. Q., Brundin, P. and Song, J. (2012), Neuronal Properties, In Vivo Effects, and Pathology of a Huntington's Disease Patient-Derived Induced Pluripotent Stem Cells. STEM CELLS, 30: 2054–2062. doi: 10.1002/stem.1135
Author contributions: I.J. and N.L.: conception and design, collection and assembly of data, and data analysis and interpretation; J.Y.L.: conception and design, collection and assembly of data, data analysis and interpretation, and manuscript writing; I.H.P.: conception and design and provision of study material; K.S.P., S.H.S., C.C., D.J.C., J.K., S.H.O., D.A.S., J.T.D., and D.R.L.: collection and assembly of data; J.M.: statistical analysis; H.S.K., M.K., and K.S.K: data analysis and interpretation; G.Q.D.: conception and design and provision of study material; P.B.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; J.S.: conception and design, provision of study material, collection and assembly of data, data analysis and interpretation, financial support, manuscript writing, and final approval of manuscript. I.J., N.L., and J.-Y.L. contributed equally to this article.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS May 24, 2012.
- Issue published online: 20 AUG 2012
- Article first published online: 20 AUG 2012
- Accepted manuscript online: 24 MAY 2012 11:19AM EST
- Manuscript Accepted: 7 MAY 2012
- Manuscript Received: 19 APR 2011
- Seoul R&D Program. Grant Number: #10548
- Korea Food & Drug Administration. Grant Number: S-11-04-2-SJV-993-0-H
- Korea Health Technology R&D Project
- Ministry of Health & Welfare. Grant Number: A111016
- Basic Science Research Program through the National Research Foundation of Korea (NRF)
- Ministry of Education, Science and Technology. Grant Number: 2012R1A1A2006827
- Republic of Korea
- Swedish Research Council
Additional Supporting Information may be found in the online version of this article.
|SC_11-0388_sm_SuppTable1.pdf||98K||Supplementary Table 1|
|SC_11-0388_sm_SuppTable2.pdf||96K||Supplementary Table 2|
|SC_11-0388_sm_SuppTable3.pdf||121K||Supplementary Table 3|
|SC_11-0388_sm_supplFigure1.tif||2278K||Supporting Information Figure S1. Neuronal differentiation and characterization of iPSCs that are derived from a Huntington's disease patient (HD-iPSC). (A): Experimental scheme showing a step-wise differentiation procedure. (B): Neural rosette-like structures formed through co-culturing HDiPSC with PA6 stromal cells (Stage 3, see the text for details). Arrow indicates a representative neural rosette-like structure. (C): Neurospheres formed though suspension culture of isolated neural rosette-like structures (Stage 4). (D): Neural precursor cells undergoing expansion as single cells (Stage 5-NP), which were made by dissociation of neurospheres using accutase. (E): Differentiated neuronal cells formed from the attached neurospheres (Stage 5-MN).|
|SC_11-0388_sm_supplFigure2.tif||2445K||Supporting Information Figure S2. Analyses of chromosomal changes and CAG repeat number changes in HD-iPSC. (A): Karyotypic analysis showing normal chromosome morphology and numbers after extensive expansion or neuronal differentiation of HD-iPSC (same results). (B): Genomic DNA PCR analysis showing no changes of CAG repeat numbers (i.e., 72 repeats) after extensive expansion (i.e., 62 more passages from passage 11 to passage 73) of HD-iPS cells. Arrow indicates the mutant huntingtin allele. No additional band was detected from hESC after extensive expansion (i.e., 73 more passages from passage 36 to passage 109) or normal control. CAG repeat numbers were counted by sequencing. (C): Genomic DNA PCR analysis showing no changes of CAG repeat numbers after neuronal differentiation of HD-iPSC. Arrow indicates the mutant huntingtin allele. No additional band was detected after neuronal differentiation of hESC. CAG repeat numbers were counted as above. Note that HD gene is highly polymorphic and father and mother may carry different numbers of CAG repeats, which will result in different sizes of PCR bands, as shown in (B) and (C).|
|SC_11-0388_sm_supplFigure3.tif||595K||Supporting Information Figure S3. Comparison of neuronal differentiation efficiency in H9, F5, HD and HD2 cells, in conjunction with transgene expression. (A): Histogram showing the percentage of rosette-forming colonies out of total colonies at Stage 3. (B): Percentage of neural forming area in each colony at Stage 3. (C): Real-time qPCR results showing the relative endogenous mRNA expression levels of four Yamanaka factors (Oct4, Sox2, Klf4 and c-Myc) in H9, F5, HD and HD2 cells. Expression levels of each gene were adjusted to those in H9 cells. Note the relatively strong expression of Klf4 gene in F5 and HD2 cells. (D): Real-time qPCR results showing the relative expression levels of Sox2 transgene at Stage 5-NP. Results are shown in a log scale. Note the low level of Sox2 transgene expression in HD cells.|
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