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Article first published online: 15 MAY 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 6, pages 1134–1143, June 2012
How to Cite
Mohanty, S. T., Cairney, C. J., Chantry, A. D., Madan, S., Fernandes, J. A., Howe, S. J., Moore, H. D., Thompson, M. J., Chen, B., Thrasher, A., Keith, W. N. and Bellantuono, I. (2012), A Small Molecule Modulator of Prion Protein Increases Human Mesenchymal Stem Cell Lifespan, Ex Vivo Expansion, and Engraftment to Bone Marrow in NOD/SCID Mice. STEM CELLS, 30: 1134–1143. doi: 10.1002/stem.1065
Author contributions: S.T.M.: collection and assembly of data, data analysis, writing of the manuscript and final approval; C.J.C.: collection and assembly of data, data analysis and interpretation, final approval of manuscript; A.D.C., S.M., and J.F.S.J.H.: conception and design, provision of study material, final approval, H.D.M.: conception and design of the project, M.J.T.: synthesis of compound 3000689(3/689), B.C.: conception, design of the project, design of indole-3-glyoxylamide family of antiprion compounds, A.T.: conception and design, provision of study material, final approval; W.N.K.: conception and design, data analysis 2 and interpretation, final approval of manuscript; I.B.: conception and design, data analysis and interpretation, manuscript writing and final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS February 24, 2012.
- Issue published online: 15 MAY 2012
- Article first published online: 15 MAY 2012
- Accepted manuscript online: 24 FEB 2012 11:07AM EST
- Manuscript Accepted: 26 JAN 2012
- Manuscript Received: 18 NOV 2011
- Biotechnology and Biological Sciences Research Council (BBSRC, BB/E014119/1)
Additional Supporting Information may be found in the online version of this article.
|SC_11-1097_sm_supplFigure1.tif||1939K||Fig. S1 hMSC shows signs of osteogenic and adipogenic differentiation. hMSC were exposed to osteogenic differentiation supplements (A-B) or adipogenic differentiation supplements (C-D) for 14 days and assessed for expression of osteogenic markers (A) osteocalcin and (B) alkaline phosphatase and adipogenic markers (C) PPAR-γ and (D) Oil red O. UD, undifferentiated; OB, exposed to osteogenic differentiation supplements; AD, exposed to adipogenic differentiation supplements.|
|SC_11-1097_sm_supplFigure2.tif||1142K||Fig. S2 Knock down of PrP expression inhibits differentiation of hMSC cultures. Realtime qPCR of untransduced (UT) hMSC cultures or transduced with shRNA-2 (sh-2) , ShRNA-ns (sh-ns), not exposed (d0) and exposed to osteogenic and adipogenic differentiation supplements for 14 days (d14) and assessed for the expression of osteogenic differentiation marker (A) CBFA-1, (B) alkaline phosphatase (ALP), (C) osteocalcin (OC) and (D) osteopontin (OP) and adipogenic differentiation marker (E) peroxisome proliferatoractivated receptor γ (PPAR-γ) and (F) Lipoprotein lipase (LPL). All markers were normalised to ribosomal protein L-32. All data are presented as mean ± SEM and analysed by one way Anova and Bonferroni multiple comparison post test with *p<0.05, **p<0.01, ***P<0.001.|
|SC_11-1097_sm_supplFigure3.tif||1112K||Fig. S3 3/689 enhances differentiation of hMSC cultures to the osteogenic lineage. Realtime qPCR of hMSC cultures expanded in medium alone (NT) or in presence of either 3/689 or DMSO prior to the exposure to osteogenic and adipogenic differentiation supplements for 14 days. At the end of this period cultures were assessed for the expression of osteogenic differentiation markers (A) CBFA-1, (B) alkaline phosphatase (ALP), (C) osteocalcin (OC) and (D) osteopontin (OP), and adipogenic differentiation markers (E) peroxisome proliferator-activated receptor γ (PPAR-γ) and (F) Lipoprotein lipase (LPL). All markers were normalised to ribosomal protein L-32. All data are presented as mean ± SEM and analysed by Bonferroni multiple comparison test with *p<0.05, **p<0.01, ***P<0.001|
|SC_11-1097_sm_supplFigure4.pdf||202K||Fig. S4. Representative examples of cells immunostained for eGFP expression considered pericyte (i), stromal cells (ii), bone lining cells (iii) and osteocyte (iv) based on morphology and anatomical location. Cells were scored 5 weeks after transplantation of eGFP+ hMSC in the bone marrow of NOD/SCID mice. Images were taken using a light microscope Leica Leits DMRB at 20× magnification.|
|SC_11-1097_sm_supplFigure5.tif||2972K||Fig. S5 3/689 abrogates dysregulation of networks related to DNA damage and cell cycle Graphical representation of the proportion of biological networks grouped according to their function and containing a significant number of differentially expressed genes, obtained by comparing gene expression profiling of hMSC cultures at passage 2 and 8 cultured in absence (A) or presence of 3/689 (B).|
|SC_11-1097_sm_supplTable1.pdf||72K||Supplementary Table 1.|
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