Thermotropic Liquid Crystals as Substrates for Imaging the Reorganization of Matrigel by Human Embryonic Stem Cells

We have investigated the culture of human embryonic stem cells (hESCs) on interfaces of the thermotropic liquid crystal, TL205, that are decorated with thin films of the extracellular matrix, Matrigel. hESCs seeded at the liquid-crystal/Matrigel interface survive for weeks, and cell colonies grow over this time. The cells show levels of differentiation comparable to that observed for cells on Matrigel-coated glass controls. Polarized and fluorescence microscopy reveal that the orientational order of the liquid crystal is coupled to the presence and organization of Matrigel. This enables straightforward imaging of the reorganization of Matrigel by the hESCs through changes in the appearance of the liquid crystal when observed using polarized light microscopy. The coupling between Matrigel and TL205 thus provides a simple tool for monitoring the reorganization of the Matrigel film over time. Our results suggest new approaches to the culture of cells and measurements of cell–extracellular-matrix interactions.

[1]  Ralph Müller,et al.  Synthetic extracellular matrices for in situ tissue engineering , 2004, Biotechnology and bioengineering.

[2]  B Jerome,et al.  Surface effects and anchoring in liquid crystals , 1991 .

[3]  N. A. Kefalides,et al.  Studies on human laminin and laminin-collagen complexes. , 1991, Connective tissue research.

[4]  Punit Ahluwalia,et al.  Integrated lithographic membranes and surface adhesion chemistry for three-dimensional cellular stimulation. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[5]  J. Goodby Liquid crystals and life , 1998 .

[6]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[7]  Martin Caffrey,et al.  Membrane protein crystallization. , 2003, Journal of structural biology.

[8]  Y. Wang,et al.  Cell locomotion and focal adhesions are regulated by substrate flexibility. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[9]  A. Harris,et al.  Silicone rubber substrata: a new wrinkle in the study of cell locomotion. , 1980, Science.

[10]  M. Dembo,et al.  Traction force microscopy of migrating normal and H-ras transformed 3T3 fibroblasts. , 2001, Biophysical journal.

[11]  N. Abbott,et al.  Formation and characterization of phospholipid monolayers spontaneously assembled at interfaces between aqueous phases and thermotropic liquid crystals. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[12]  C. Murphy,et al.  Adhesion and proliferation of corneal epithelial cells on self-assembled monolayers. , 2000, Journal of biomedical materials research.

[13]  B. G. Anex Optical Properties of Highly Absorbing Crystals , 1966 .

[14]  L. Liotta,et al.  Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. , 1982, Biochemistry.

[15]  P. Janmey,et al.  Biomechanics and Mechanotransduction in Cells and Tissues Cell type-specific response to growth on soft materials , 2005 .

[16]  Juan J de Pablo,et al.  Inhibition of human embryonic stem cell differentiation by mechanical strain , 2006, Journal of cellular physiology.

[17]  J. Wong,et al.  Proliferation of dinoflagellates: blooming or bleaching , 2005, BioEssays : news and reviews in molecular, cellular and developmental biology.

[18]  Christopher S. Chen,et al.  Cells lying on a bed of microneedles: An approach to isolate mechanical force , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[19]  C. S. Chen,et al.  Geometric control of cell life and death. , 1997, Science.

[20]  K. Nakano Scaling Law on Molecular Orientation and Effective Viscosity of Liquid-Crystalline Boundary Films , 2003 .

[21]  N. Abbott,et al.  Biomolecular Interactions at Phospholipid-Decorated Surfaces of Liquid Crystals , 2003, Science.

[22]  D. Kaufman,et al.  Multilineage Differentiation from Human Embryonic Stem Cell Lines , 2001, Stem cells.

[23]  F M Watt,et al.  Out of Eden: stem cells and their niches. , 2000, Science.

[24]  V. Norris,et al.  Chromosome separation and segregation in dinoflagellates and bacteria may depend on liquid crystalline states. , 2001, Biochimie.

[25]  A. Engler,et al.  Photopolymerization in Microfluidic Gradient Generators: Microscale Control of Substrate Compliance to Manipulate Cell Response , 2004 .

[26]  Dennis Discher,et al.  Substrate compliance versus ligand density in cell on gel responses. , 2004, Biophysical journal.

[27]  M. Sheetz,et al.  Forces on adhesive contacts affect cell function. , 1998, Current opinion in cell biology.

[28]  Benjamin G Keselowsky,et al.  Surface chemistry modulates focal adhesion composition and signaling through changes in integrin binding. , 2004, Biomaterials.

[29]  C. Streuli,et al.  Extracellular matrix remodelling and cellular differentiation. , 1999, Current opinion in cell biology.

[30]  W. Saltzman,et al.  Topographical control of human neutrophil motility on micropatterned materials with various surface chemistry. , 2002, Biomaterials.

[31]  C. Frank,et al.  Binary self-assembled monolayers: spectroscopy and application to liquid crystal alignment , 1994 .

[32]  Joyce Y. Wong,et al.  Balance of chemistry, topography, and mechanics at the cell–biomaterial interface: Issues and challenges for assessing the role of substrate mechanics on cell response , 2004 .

[33]  L. Smeller Pressure-temperature phase diagrams of biomolecules. , 2002, Biochimica et biophysica acta.

[34]  N. Abbott,et al.  An Experimental System for Imaging the Reversible Adsorption of Amphiphiles at Aqueous−Liquid Crystal Interfaces , 2002 .

[35]  C. Murphy,et al.  Non-toxic thermotropic liquid crystals for use with mammalian cells , 2004 .

[36]  Ranganathan Shashidhar,et al.  Imaging Biological Cells Using Liquid Crystals , 2003 .