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71 Eyckmans, J. et al., 2011. A hitchhiker’s guide to mechanobiology. Developmental cell, 21(1), pp.35–47. Available at: http://www.ncbi.nlm.nih.gov/pubmed/21763607 [Accessed March 8, 2012]. Friedenstein, A.J. et al. (1974). Precursors for fibroblasts in different populations of hematopoietic cells as detected by the in vitro colony assay method. Experimental hematology 2, 83-92. Goh Jih Her, Hsi-Chin Wu, Ming-Hong Chen, Ming-Yi Chen, Shun-Chih Chang, Tzu-Wei Wan. Control of three-dimensional substrate stiffness to manipulate mesenchymal stem cell fate toward neuronal or glial lineages. Acta Biomaterialia 9 (2013) 5170–5180 Hao, H., Chen, G., Liu, J., Ti, D., Zhao, Y., Xu, S., … Han, W. (2013). Culturing on Wharton’s Jelly Extract Delays Mesenchymal Stem Cell Senescence through p53 and p16INK4a/pRb Pathways. PloS one, 8(3), e58314. doi:10.1371/journal.pone.0058314 Her, G. J., Wu, H.-C., Chen, M.-H., Chen, M.-Y., Chang, S.-C., & Wang, T.-W. (2013). Control of three-dimensional ssubtrate stiffness to manipulate mesenchymal stem cell fate toward neuronal or glial lineages. Acta biomaterialia, 9(2), 5170–80. doi:10.1016/j.actbio.2012.10.012 Hermann, A. et al. Comparative analysis of neuroectodermal differentiation capacity of human bone marrow stromal cells using various conversion protocols. Journal of neuroscience research 83, 1502-14 (2006). Hoffecker, I. T., Guo, W.-h. & Wang, Y.-l. (2011). Assessing the spatial resolution of cellular rigidity sensing using a micropatterned hydrogel–photoresist composite. Lab on a Chip 11, 3538, doi:10.1039/c1lc20504h. Huang, S., Chen, C.S. & Ingber, D.E., 1998. Control of cyclin D1, p27(Kip1), and cell cycle progression in human capillary endothelial cells by cell shape and cytoskeletal tension. Molecular biology of the cell, 9(11), pp.3179–93. Hung, H.-S., Tang, C.-M., Lin, C.-H., Lin, S.-Z., Chu, M.-Y., Sun, W.-S., … Hsu, S. (2013). Biocompatibility and favorable response of mesenchymal stem cells on fibronectingold nanocomposites. PloS one, 8(6), e65738. doi:10.1371/journal.pone.0065738 Katsetos CD, Herman MM, Mörk SJ. Class III b-tubulin in human development and cancer. Cell Motil Cytoskeleton 2003;55:77–96. Kippert, A., Fitzner, D., Helenius, J. & Simons, M. (2009). Actomyosin contractility controls cell surface area of oligodendrocytes. BMC Cell Biology 10, 71, doi:10.1186/1471-2121-10-71 Lourenço, T. (2012). Regulation of ECM mimetics during oligodendrocytes differentiation in vitro. In Life Science (Coimbra: University of Coimbra)
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DEPARTAMENTO DE CIÊNCIAS DA VIDA F
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ii Queria agradecer à minha namora
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iv isolation of hUCM-MSCs was ever
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vi Conseguimos optimizar um novo pr
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viii Table of contents Acknowledgem
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x III. 6 - Influence on MSCs specif
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xii Figure 2| Proteins related to t
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xiv the change in distribution with
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xvi Cells were stained with anti-B-
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xviii List of abbreviations AA - Ac
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1 Chapter I Introduction
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4 There are many advantages in usin
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6 only ones MSCs can differentiate
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Figure 1. Mechanotransduction in a
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10 1C)(Dogterom et al., 2005). Not
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12 extracellular ligand and the cyt
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14 Figure 3 - The Rigidity Sensing
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16 Figure 5 - Images depicted on A)
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18 Figure 7 - A) Immunofluorescence
Page 41 and 42: 20 (GFAP) (is a class-III intermedi
Page 43 and 44: 22 can specify lineage towards cell
Page 45 and 46: 24 I.3 - Project rationale and expe
Page 47 and 48: 26 Figure 12 - MSCs cultured on 1 a
Page 49 and 50: 28 than that in soft substrate. Thi
Page 52: 31 Chapter II - Materials and Metho
Page 55 and 56: 34 Covance and DakoCytomation, resp
Page 57 and 58: 36 clone SAM1; mouse Pe-Cy7 anti-hu
Page 59 and 60: 38 To functionalize the complete su
Page 62: 41 Chapter III Results
Page 65 and 66: 44 2008) and also as previously est
Page 67 and 68: 12.5% Ac PBS Fold increase of hMSCs
Page 69 and 70: ≈12 kPa / COL-1 +FN ≈12 kPa / C
Page 71 and 72: 50 Figure 19 - Immunophenotype of U
Page 73 and 74: 52 coated with COL-1 (similar to wh
Page 75 and 76: 54 hydrogels of 1 and 7 kPa (for ce
Page 77: Figure 23 - hMSCs cultured on TCP (
Page 81 and 82: 60 Discussion MSCs are widely used
Page 83 and 84: 62 isolating MSCs on the PA hydroge
Page 85 and 86: 64 2011) and expanded on TCPs for 5
Page 88: 67 Conclusion In conclusion, we opt
Page 91: 70 Cretu, A., Castagnino, P. & Asso
Page 95: 74 Wang, N., Butler, J.P., and Ingb
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