BE 167L Lecture 6 General

BE 167L Lecture 6 General - BE 167L – Bioengineering...

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Unformatted text preview: BE 167L – Bioengineering Lab Lecture 6 Today •  2D vs. 3D Cell Culture and the cell microenvironment – 2D Culture – 3D Culture – Literature examples •  3D encapsulaCon for lab next week Tuesday Mammary Branching Morphogenesis Case Study Why 2D / 3D Cultures? 2D Cultures •  Simple to grow and feed cells •  Easy to image cells with microscopy 3D Cultures •  Understanding cell behavior with in vivo relevance •  Develop scaffolds for Cssue engineering •  Create environments to differenCate stem cells to desired lineages Lippincott-Schwartz Lab Input ­Output Response of a Cell 2D 3D 3D Yarmush & King. Annual Review Biomed Engineering 2009 The Cell Microenvironment (2D) Soluble Factors •  Cytokines •  Growth Factors Interleukin-8 (IL-8) •  Hormones •  Nutrients Epinephrine Epidermal Growth Factor (EGF) Cell ­ECM Contact ECM – Extracellular Matrix Protein Components: Laminin, Collagen, Elastin, Fibronectin Proteoglycan Components: Heparan Sulfate, Chondroitin Sulfate Sugar Components: Hyaluronic Acid Cell ­Cell CommunicaCon •  Gap JuncCons – connexons – < 1kD molecules can pass through •  Adherent JuncCons – cadherins Pa\erned 2D Cultures to Study ECM, Cell Shape, and Cell Contacts Geometric Control of Cell Life and Death Chen et al. Science 1997, vol 276 Controlling for ECM concentraCon Chen et al. Science 1997, vol 276 Lumen FormaCon on Pa\erned Surfaces Is this believable? Dike et al. In Vitro Cellular and Developmental Biology 3D Cultures for Studying Tissue-like Behavior Stelzer et al. Nat Rev Mol Cell Biol 2007 Controlling Cell ­Cell and Cell ­ECM Contact Todd McDevitt, Georgia Tech •  Spheroids / Colonies – Embryoid bodies – Hepatocyte spheroids – Cancer spheroids Embryoid Bodies (EBs) •  3D Matrix – Hydrogels – Others Spheroid Cultures Key Concepts: Unique Cell-Cell Contact Environment Unique Mass Transport Environment Gels for Cell EncapsulaCon •  Immune isolaCon – Xenogra_s – Diabetes, hemophilia, cancer, renal failure •  RegeneraCve medicine – Stem cells, progenitor cells 17 Orive et al. Nature Medicine 9, 104 - 107 (2003) 3D Cell Culture for Tissue Engineering Khademhosseini et al. PNAS 2006 Alginates: examples of ionic gels 2nd most abundant biopolymer  Anionic polysaccharides from seaweeds     Glucoronic & Mannuronic acids Reacts with Ca2+ ions to form gels   19 General ClassificaCons of Gels in Tissue Engineering •  Chemical gels – Primary bonds (covalent) – pH, ionic groups, solvent concentraCon – Usually irreversible •  Physical gels – Secondary bonds – temperature, shear, viscosity – Usually reversible 20 “The Encapsulator ” 21 Your Encapsulator Cells in alginate gel CaCl2 22 RoboCc cell wriCng 23 Rapid Prototyping Journal 2005: 11(1) 9-17 RoboCc cell wriCng 24 Rapid Prototyping Journal 2005: 11(1) 9-17 PDMS based cell encapsulaCon 25 Qiu et al. Adv. Mat 2007, 19, 1603-7 Microfluidic based single cell encapsulaCon 26 Life beyond gels – Cells in other 3D systems Particulate Leaching Electrospun Fibers 27 Cell Mechanics and the Microenvironment Mini ­project Ideas, starts a_er lab pracCcal •  Test fidelity of laser cu\er mold to cell pa\erning at different sizes •  InvesCgate cell viability in different pH mediums or different soluCons •  Test cell migraCon speed across different distances •  Test viability of cells encapsulated at different densiCes ...
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This note was uploaded on 11/10/2011 for the course BE 167 taught by Professor Staff during the Spring '10 term at UCLA.

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