Electrotaxis of lung cancer cells in ordered three-dimensional scaffolds.

In this paper, we report a new method to incorporate 3D scaffold with electrotaxis measurement in the microfluidic device. The electrotactic response of lung cancer cells in the 3D foam scaffolds which resemble the in vivo pulmonary alveoli may give more insight on cellular behaviors in vivo. The 3D scaffold consists of ordered arrays of uniform spherical pores in gelatin. We found that cell morphology in the 3D scaffold was different from that in 2D substrate. Next, we applied a direct current electric field (EF) of 338 mV/mm through the scaffold for the study of cells' migration within. We measured the migration directedness and speed of different lung cancer cell lines, CL1-0, CL1-5, and A549, and compared with those examined in 2D gelatin-coated and bare substrates. The migration direction is the same for all conditions but there are clear differences in cell morphology, directedness, and migration speed under EF. Our results demonstrate cell migration under EF is different in 2D and 3D environments and possibly due to different cell morphology and/or substrate stiffness.

[1]  Grace N Li,et al.  Genomic and morphological changes of neuroblastoma cells in response to three-dimensional matrices. , 2007, Tissue engineering.

[2]  R. Isseroff,et al.  Imposition of a physiologic DC electric field alters the migratory response of human keratinocytes on extracellular matrix molecules. , 1996, The Journal of investigative dermatology.

[3]  Meng-Hua Yen,et al.  Electrotaxis of lung cancer cells in a multiple-electric-field chip. , 2009, Biosensors & bioelectronics.

[4]  Douglas A Lauffenburger,et al.  Marrow‐Derived stem cell motility in 3D synthetic scaffold is governed by geometry along with adhesivity and stiffness , 2010, Biotechnology and bioengineering.

[5]  Mechthild Krause,et al.  Three-dimensional cell growth confers radioresistance by chromatin density modification. , 2010, Cancer research.

[6]  P. Janmey,et al.  Tissue Cells Feel and Respond to the Stiffness of Their Substrate , 2005, Science.

[7]  Min Zhao,et al.  EGF receptor signalling is essential for electric-field-directed migration of breast cancer cells , 2007, Journal of Cell Science.

[8]  Shang-Tian Yang,et al.  Effects of Three‐Dimensional Culturing in a Fibrous Matrix on Cell Cycle, Apoptosis, and MAb Production by Hybridoma Cells , 2008, Biotechnology progress.

[9]  Ronald A. Li,et al.  Electrically Guiding Migration of Human Induced Pluripotent Stem Cells , 2010, Stem Cell Reviews and Reports.

[10]  Francis Lin,et al.  Lymphocyte Electrotaxis In Vitro and In Vivo1 , 2008, The Journal of Immunology.

[11]  D. Thomson,et al.  Activated T lymphocytes migrate toward the cathode of DC electric fields in microfluidic devices. , 2011, Lab on a chip.

[12]  Danny Baranes,et al.  Superior survival and durability of neurons and astrocytes on 3-dimensional aragonite biomatrices. , 2007, Tissue engineering.

[13]  John Kolega,et al.  Effects of Direct Current Electric Fields on Cell Migration and Actin Filament Distribution in Bovine Vascular Endothelial Cells , 2002, Journal of Vascular Research.

[14]  C. M. Cheah,et al.  Influence of process parameters on stereolithography part shrinkage , 1996 .

[15]  Thierry Balaguer,et al.  Human Primary Osteocyte Differentiation in a 3D Culture System , 2009, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[16]  H. K. Soong,et al.  Galvanotropic and galvanotaxic responses of corneal endothelial cells. , 1996, Journal of the Formosan Medical Association = Taiwan yi zhi.

[17]  Dietmar W Hutmacher,et al.  Combining electrospun scaffolds with electrosprayed hydrogels leads to three-dimensional cellularization of hybrid constructs. , 2008, Biomacromolecules.

[18]  Yih-Leong Chang,et al.  Collapsin response mediator protein-1 and the invasion and metastasis of cancer cells. , 2001, Journal of the National Cancer Institute.

[19]  Matsuhiko Nishizawa,et al.  Multi-channel 3-D cell culture device integrated on a silicon chip for anticancer drug sensitivity test. , 2005, Biomaterials.

[20]  Edyta Brzóska,et al.  Comparison of satellite cell‐derived myoblasts and C2C12 differentiation in two‐ and three‐dimensional cultures: changes in adhesion protein expression , 2011, Cell biology international.

[21]  I. Zein,et al.  Fused deposition modeling of novel scaffold architectures for tissue engineering applications. , 2002, Biomaterials.

[22]  S. Hui,et al.  A calcium requirement for electric field-induced cell shape changes and preferential orientation. , 1985, Cell calcium.

[23]  Shang-Tian Yang,et al.  Effects of Three‐Dimensional Culturing on Osteosarcoma Cells Grown in a Fibrous Matrix: Analyses of Cell Morphology, Cell Cycle, and Apoptosis , 2003, Biotechnology progress.

[24]  A. Godwin,et al.  Ovarian normal and tumor-associated fibroblasts retain in vivo stromal characteristics in a 3-D matrix-dependent manner. , 2008, Gynecologic oncology.

[25]  J. Elisseeff,et al.  Experimental model for cartilage tissue engineering to regenerate the zonal organization of articular cartilage. , 2003, Osteoarthritis and cartilage.

[26]  J. Shih,et al.  Global analysis of gene expression in invasion by a lung cancer model. , 2001, Cancer research.

[27]  Min Zhao,et al.  Controlling cell behavior electrically: current views and future potential. , 2005, Physiological reviews.

[28]  M. Cima,et al.  Mechanical properties of dense polylactic acid structures fabricated by three dimensional printing. , 1996, Journal of biomaterials science. Polymer edition.

[29]  Min Zhao,et al.  Electrical stimulation directly induces pre-angiogenic responses in vascular endothelial cells by signaling through VEGF receptors , 2003, Journal of Cell Science.

[30]  A. Knox,et al.  Use of a three-dimensional cell culture model to study airway smooth muscle-mast cell interactions in airway remodeling. , 2009, American journal of physiology. Lung cellular and molecular physiology.

[31]  Biman B Mandal,et al.  Cell proliferation and migration in silk fibroin 3D scaffolds. , 2009, Biomaterials.

[32]  Krishnendu Roy,et al.  Biomimetic three-dimensional cultures significantly increase hematopoietic differentiation efficacy of embryonic stem cells. , 2005, Tissue engineering.

[33]  R. Nuccitelli,et al.  Endogenous electric fields in embryos during development, regeneration and wound healing. , 2003, Radiation protection dosimetry.

[34]  Jun Li,et al.  Poly(lactic acid) scaffold fabricated by gelatin particle leaching has good biocompatibility for chondrogenesis , 2008, Journal of biomaterials science. Polymer edition.

[35]  Dietmar W. Hutmacher,et al.  Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells. , 2010, Biomaterials.

[36]  Shelly R. Peyton,et al.  The use of poly(ethylene glycol) hydrogels to investigate the impact of ECM chemistry and mechanics on smooth muscle cells. , 2006, Biomaterials.

[37]  Xiaolong Yan,et al.  Lung cancer A549 cells migrate directionally in DC electric fields with polarized and activated EGFRs , 2009, Bioelectromagnetics.

[38]  S. Rizzi,et al.  Elucidating the role of matrix stiffness in 3D cell migration and remodeling. , 2011, Biophysical journal.

[39]  David W. Holman,et al.  Characterization of arachnoidal cells cultured on three-dimensional nonwoven PET matrix. , 2007, Tissue engineering.

[40]  Stephanie H. Nowotarski,et al.  Morphology and organization of tissue cells in 3D microenvironment of monodisperse foam scaffolds , 2011 .

[41]  T. Meyer,et al.  A new method for the 3‐D in vitro growth of human RT112bladder carcinoma cells using the alginate culture technique , 1994, Biology of the cell.

[42]  Tetsuya Horiuchi,et al.  E-cadherin-dependent intercellular adhesion enhances chemoresistance. , 2003, International journal of molecular medicine.

[43]  Toshiomi Yoshida,et al.  Effect of heparin addition on expansion of cord blood hematopoietic progenitor cells in three-dimensional coculture with stromal cells in nonwoven fabrics , 2005, Journal of Artificial Organs.

[44]  M. Levin.  Bioelectric mechanisms in regeneration: Unique aspects and future perspectives. , 2009, Seminars in cell & developmental biology.

[45]  H. Gruler,et al.  Galvanotaxis of human granulocytes , 1988, European Biophysics Journal.

[46]  Z. Madeja,et al.  Directional movement of rat prostate cancer cells in direct-current electric field: involvement of voltagegated Na+ channel activity. , 2001, Journal of cell science.

[47]  A. Goldberg,et al.  TNF‐α increases ubiquitin‐conjugating activity in skeletal muscle by up‐regulating UbcH2/E220k , 2003 .

[48]  Y. Zhao,et al.  A packed Cytodex microbead array for three-dimensional cell-based biosensing. , 2006, Biosensors & bioelectronics.

[49]  E. Bois-Reymond.  Untersuchungen über thierische Elektricität , 1848 .

[50]  S. MacNeil,et al.  A novel route for the production of chitosan/poly(lactide-co-glycolide) graft copolymers for electrospinning , 2010, Biomedical materials.

[51]  Tao Tian,et al.  Survival advantages of multicellular spheroids vs. monolayers of HepG2 cells in vitro. , 2008, Oncology reports.

[52]  M. S. Cooper,et al.  Perpendicular orientation and directional migration of amphibian neural crest cells in dc electrical fields. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[53]  D. Ornstein,et al.  The increased expression of periostin during early stages of prostate cancer and advanced stages of cancer stroma , 2009, The Prostate.

[54]  Min Zhao,et al.  Application of direct current electric fields to cells and tissues in vitro and modulation of wound electric field in vivo , 2006, Nature Protocols.

[55]  Q. Meng.  Three-dimensional culture of hepatocytes for prediction of drug-induced hepatotoxicity , 2010, Expert opinion on drug metabolism & toxicology.

[56]  Maria Bokhari,et al.  Culture of HepG2 liver cells on three dimensional polystyrene scaffolds enhances cell structure and function during toxicological challenge , 2007, Journal of anatomy.

[57]  B. Heng,et al.  Comparison of osteogenesis of human embryonic stem cells within 2D and 3D culture systems , 2008, Scandinavian journal of clinical and laboratory investigation.

[58]  S. Sahoo,et al.  3-D tumor model for in vitro evaluation of anticancer drugs. , 2008, Molecular pharmaceutics.

[59]  Itai Cohen,et al.  Mapping the depth dependence of shear properties in articular cartilage. , 2007, Journal of biomechanics.

[60]  Biaoyang Lin,et al.  A porous 3D cell culture micro device for cell migration study , 2010, Biomedical microdevices.

[61]  A. Khademhosseini,et al.  Micro‐Masonry: Construction of 3D Structures by Microscale Self‐Assembly , 2010, Advanced materials.

[62]  Charles Keith,et al.  Human neuroblastoma (SH-SY5Y) cell culture and differentiation in 3-D collagen hydrogels for cell-based biosensing. , 2006, Biosensors & bioelectronics.

[63]  Keng-hui Lin,et al.  Fabricating scaffolds by microfluidics. , 2009, Biomicrofluidics.

[64]  M. Hendrix,et al.  Selection of invasive and metastatic subpopulations from a human lung adenocarcinoma cell line. , 1997, American journal of respiratory cell and molecular biology.

[65]  D. Kniss,et al.  Three-dimensional cell-scaffold constructs promote efficient gene transfection: implications for cell-based gene therapy. , 2001, Tissue engineering.

[66]  Yen Wei,et al.  Electrospinning polyaniline-contained gelatin nanofibers for tissue engineering applications. , 2006, Biomaterials.

[67]  C. Chen,et al.  Preparation and Properties of Poly(lactide-co-glycolide) (PLGA)/ Nano-Hydroxyapatite (NHA) Scaffolds by Thermally Induced Phase Separation and Rabbit MSCs Culture on Scaffolds , 2008, Journal of biomaterials applications.

[68]  Tessa Lühmann,et al.  The induction of cell alignment by covalently immobilized gradients of the 6th Ig-like domain of cell adhesion molecule L1 in 3D-fibrin matrices. , 2009, Biomaterials.

[69]  Yonghong Sun,et al.  Differential methotrexate hepatotoxicity on rat hepatocytes in 2-D monolayer culture and 3-D gel entrapment culture. , 2009, Chemico-biological interactions.

[70]  Min Zhao,et al.  Electrotaxis and wound healing: experimental methods to study electric fields as a directional signal for cell migration. , 2009, Methods in molecular biology.

[71]  Stephanie I. Fraley,et al.  A distinctive role for focal adhesion proteins in three-dimensional cell motility , 2010, Nature Cell Biology.

[72]  Y. H. Park,et al.  Development of 3-D nanofibrous fibroin scaffold with high porosity by electrospinning: implications for bone regeneration , 2008, Biotechnology Letters.