Telephone: 905-525-9140, ext. 28687; Fax: 905-522-7772
Embryonic Stem Cells/Induced Pluripotent Stem Cells
Version of Record online: 18 JAN 2012
Copyright © 2011 AlphaMed Press
Volume 30, Issue 2, pages 131–139, February 2012
How to Cite
Risueño, R. M., Sachlos, E., Lee, J.-H., Lee, J. B., Hong, S.-H., Szabo, E. and Bhatia, M. (2012), Inability of Human Induced Pluripotent Stem Cell-Hematopoietic Derivatives to Downregulate MicroRNAs In Vivo Reveals a Block in Xenograft Hematopoietic Regeneration. STEM CELLS, 30: 131–139. doi: 10.1002/stem.1684
Author contributions: R.M.R.: conception and design, collection and/or assembly of data, data analysis and interpretation, and manuscript writing, E.S.: conception and design, collection and/or assembly of data, data analysis and interpretation, and manuscript writing, J.-H.L., J.B.L., S.-H.H, and E. Szabo: collection and/or assembly of data and data analysis and interpretation; M.B.: conception and design, financial support, data analysis and interpretation, manuscript writing, and final approval of the manuscript. R.M.R. and E.S. 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 November 30, 2011.
- Issue online: 18 JAN 2012
- Version of Record online: 18 JAN 2012
- Accepted manuscript online: 30 NOV 2011 01:47PM EST
- Manuscript Accepted: 11 NOV 2011
- Manuscript Received: 11 AUG 2011
- Canada Research Chair in human stem cell biology
- TTF fellowship (CCSRI)
- Ministry of Research and Innovation fellowship
Additional Supporting Information may be found in the online version of this article.
|STEM_1684_sm_supptable1.pdf||15K||Table S1, related to Fig 3: Candidate genes relevant to the various processes of hematopoietic reconstitution. List of genes relevant in each stage of hematopoietic reconstitution used in cluster analysis Fig S5.|
|STEM_1684_sm_supptable2.pdf||48K||Table S2: Changes in candidate miRNA levels following reconstitution. List of changes in miRNA expression during engraftment of HSC in xenotransplantation with miRNAs being either A. downregulated or turned off versus B. miRNAs becoming detected i.e. turned ON.|
|STEM_1684_sm_supptable3.pdf||17K||Table S2. Top 20 ranked targets for each miRNA candidate.|
|STEM_1684_sm_supplFig1.pdf||197K||Figure S1, related to Fig 1: Characterization of iPSCs and analysis of pluripotency. A. Reprogramming of fibroblasts to iPSCs induced a transition to hESC-like colony morphology. B. Flow cytometry analysis of hESCs and iPSCs for pluripotency markers including Oct4, Tra 1-81, SSEA3 and SSEA4. C. Intratesticular delivery of iPSCs in immuno-deficient mouse hosts generated teratomas, which display all three germ layers. D. Hematopoietic development of two iPSC lines derived from human fibroblasts relative to H9 hESC. Scale bar, 500μm. E. Quantitative PCR showing the levels of total Lin28 of hiPSC relative H9 hESC confirming silencing of this reprogramming factor.|
|STEM_1684_sm_supplFig2.pdf||71K||Figure S2, related to Fig 2: Engraftment of human cells in xenotransplantation models. A. Adult HSCs readily engrafted NOD-SCID recipients and can be detected in the injected femur and the contralateral bones, unlike hESC or iPSCs. B. Whole gel PCR showing human specific Chromosome 17 expression in the injected femur (F), contralateral bones (C) or spleen (S) of NSG mice transplanted with HSC, hESC-derived or hiPSC-derived cells.|
|STEM_1684_sm_supplFig3.pdf||160K||Figure S3, related to Fig 2: Analysis of hiPSC-graft cells. A. iPSC-derived graft cells were also devoid of several lineage specific markers including CD33 (myeloid), CD3 (T-cell) and CD19 (B-cell). For comparison, cells extracted from mice injected with HSCs are also presented. B. Live-dead discrimination plots of hiPSC-graft cells showing the presence of HLA-A/B/C.|
|STEM_1684_sm_supplFig4.pdf||44K||Figure S4, related to Fig 3: Image of colony forming units (CFU). CFUs generated from A. hiPSC-derived graft cells or B. HSC graft cells recovered from xenotransplantation recipients.|
|STEM_1684_sm_supplFig5.pdf||66K||Figure S5, related to Fig 3: Cluster analysis of various processes involved in hematopoietic reconstitution.|
|STEM_1684_sm_supplFig6.pdf||50K||Figure S6. related to Fig 3: miRNA heat map of hESC-derivatives. miRNA heat map of hESC-derivatives and their comparison to HSC and engrafted HSCs.|
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.