DEPARTAMENTO DE CIÊNCIAS DA VIDA ... - Estudo Geral

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15 I.2.3 - Mechanotransduction and Stem Cells I.2.3.1 - Mechanotransduction and MSCs differentiation Living tissues are known to possess different physiologic characteristics according to their function and cellular type, so considering the elasticity of solid tissues, very different values of elasticity can be found as shown in Figure 4. The solid tissues exhibit a range of stiffness, as measured by the elastic modulus, E (Engler et al., 2006). Figure 4 – Tissue Elasticity. Range of stiffness measured by the elastic modulus, E, of some human solid tissues. Adapted from Engler et al., 2006. Mesenchymal stem cells will differentiate into different phenotypes as a function of substrate stiffness. The lineage they can differentiate into when cultured on a substrate with a certain range of stiffness has phenotypic features similar to cells found in the solid tissues with the same range of stiffness (Figure 4), and MSCs appear to do so in a way that would promote tissue-specific differentiation and healing (Engler et al., 2006). For example, on brain-tissue-like stiffness cells undergo neuronal-like differentiation, whereas muscle-equivalent stiffness promotes myogenesis (Figure 5) (Engler et al., 2006).
16 Figure 5 - Images depicted on A) and B) quantify the morphological changes (mean ± SEM) versus stiffness, E: shown are (A) cell branching per length of primary mouse neurons, MSCs, and blebbistatintreated MSCs and (B) spindle morphology of MSCs, blebbistatin-treated MSCs, and mitomycin-C treated MSCs (open squares) compared to C2C12 myoblasts (dashed line). Furthermore in C) MSCs change their morphology developing increasingly branched, spindle, or polygonal shapes, respectively, when cultured on matrices with the elastic modulus (E) respectively in the range typical of brain (0.1–1 kPa), muscle (8– 17 kPa), or stiff crosslinked-collagen matrices (25–40 kPa). Blebbistatin blocks morphology changes due to stiffness (
Page 1 and 2: DEPARTAMENTO DE CIÊNCIAS DA VIDA F
Page 3 and 4: ii Queria agradecer à minha namora
Page 5 and 6: iv isolation of hUCM-MSCs was ever
Page 7 and 8: vi Conseguimos optimizar um novo pr
Page 9 and 10: viii Table of contents Acknowledgem
Page 11 and 12: x III. 6 - Influence on MSCs specif
Page 13 and 14: xii Figure 2| Proteins related to t
Page 15 and 16: xiv the change in distribution with
Page 17 and 18: xvi Cells were stained with anti-B-
Page 19 and 20: xviii List of abbreviations AA - Ac
Page 22: 1 Chapter I Introduction
Page 25 and 26: 4 There are many advantages in usin
Page 27 and 28: 6 only ones MSCs can differentiate
Page 29 and 30: Figure 1. Mechanotransduction in a
Page 31 and 32: 10 1C)(Dogterom et al., 2005). Not
Page 33 and 34: 12 extracellular ligand and the cyt
Page 35: 14 Figure 3 - The Rigidity Sensing
Page 39 and 40: 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
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67 Conclusion In conclusion, we opt
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70 Cretu, A., Castagnino, P. & Asso
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72 McBeath, R. et al., 2004. Cell s
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74 Wang, N., Butler, J.P., and Ingb
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