A simple cell patterning method using magnetic particle-containing photosensitive poly (ethylene glycol) hydrogel blocks: a technical note.

All human organs consist of multiple types of cells organized in a complex pattern to meet specific functional needs. One possible approach for reconstructing human organs in vitro is to generate cell sheets of a specific pattern and later stack them systematically by layer into a three-dimensional organoid. However, many commonly used cell patterning techniques suffer drawbacks such as dependence on sophisticated instruments and manipulation of cells under suboptimal growth conditions. Here, we describe a simple cell patterning method that may overcome these problems. This method is based on magnetic force and photoresponsive poly (ethylene glycol) diacrylate (PEG-DA) hydrogels. The PEG-DA hydrogel was magnetized by mixing with iron ferrous microparticles and then fabricated into blocks with a specific pattern by photolithography. The resolution of the hydrogel empty space pattern was approximately 150  μm and the generated hydrogel blocks can be remotely manipulated with a magnet. The magnetic PEG-DA blocks were used as a stencil to define the area for cell adhesion in the cell culture dish, and the second types of cells could be seeded after the magnetic block was removed to create heterotypic cell patterns. Cell viability assay has demonstrated that magnetic PEG-DA and the patterning process produced negligible effects on cell growth. Together, our results indicate that this magnetic hydrogel-based cell patterning method is simple to perform and is a useful tool for tissue surrogate assembly for disease mechanism study and drug screening.

[1]  Helene Andersson,et al.  Microfabrication and microfluidics for tissue engineering: state of the art and future opportunities. , 2004, Lab on a chip.

[2]  P. Chavrier,et al.  Collective migration of an epithelial monolayer in response to a model wound , 2007, Proceedings of the National Academy of Sciences.

[3]  I. Dubery,et al.  Assessment of a simple, non-toxic Alamar blue cell survival assay to monitor tomato cell viability. , 2001, Phytochemical analysis : PCA.

[4]  Mitsuhiro Shikida,et al.  Cell culture arrays using magnetic force-based cell patterning for dynamic single cell analysis. , 2008, Lab on a chip.

[5]  Sungho Jin,et al.  Nanotoxicity of iron oxide nanoparticle internalization in growing neurons. , 2007, Biomaterials.

[6]  Masayuki Yamato,et al.  Cell micropatterning using photopolymerization with a liquid crystal device commercial projector. , 2004, Biomaterials.

[7]  J. Vacanti,et al.  Tissue engineering. , 1993, Science.

[8]  Ming C. Wu,et al.  Massively parallel manipulation of single cells and microparticles using optical images , 2005, Nature.

[9]  L. Griffith,et al.  Capturing complex 3D tissue physiology in vitro , 2006, Nature Reviews Molecular Cell Biology.

[10]  Ali Khademhosseini,et al.  Micropatterned cell co-cultures using layer-by-layer deposition of extracellular matrix components. , 2006, Biomaterials.

[11]  Hiroyuki Honda,et al.  Cell patterning using magnetite nanoparticles and magnetic force , 2007, Biotechnology and bioengineering.

[12]  Yoshinori Kawabe,et al.  Construction of heterotypic cell sheets by magnetic force-based 3-D coculture of HepG2 and NIH3T3 cells. , 2007, Journal of bioscience and bioengineering.

[13]  Hiroyuki Honda,et al.  Construction and harvest of multilayered keratinocyte sheets using magnetite nanoparticles and magnetic force. , 2004, Tissue engineering.

[14]  Masayuki Yamato,et al.  Second-generation maskless photolithography device for surface micropatterning and microfluidic channel fabrication. , 2008, Analytical chemistry.

[15]  Masayuki Yamato,et al.  Cell sheet technology and cell patterning for biofabrication , 2009, Biofabrication.

[16]  Jeroen Rouwkema,et al.  Tissue assembly and organization: developmental mechanisms in microfabricated tissues. , 2009, Biomaterials.

[17]  Cheng-Hsien Liu,et al.  Dielectrophoresis based‐cell patterning for tissue engineering , 2006, Biotechnology journal.

[18]  Masayuki Yamato,et al.  Maskless liquid-crystal-display projection photolithography for improved design flexibility of cellular micropatterns. , 2006, Biomaterials.

[19]  Xudong Cao,et al.  Patterning multiple cell types in co-cultures: A review , 2009 .

[20]  Shinji Sugiura,et al.  Stepwise assembly of micropatterned co‐cultures using photoresponsive culture surfaces and its application to hepatic tissue arrays , 2009, Biotechnology and bioengineering.

[21]  T. Okano,et al.  Cell sheet engineering: a unique nanotechnology for scaffold‐free tissue reconstruction with clinical applications in regenerative medicine , 2010, Journal of internal medicine.

[22]  M. Ozkan,et al.  Electro-optical platform for the manipulation of live cells , 2003 .

[23]  A. Ito,et al.  Fabrication of complex three-dimensional tissue architectures using a magnetic force-based cell patterning technique , 2009, Biomedical microdevices.

[24]  G. Whitesides,et al.  Patterned deposition of cells and proteins onto surfaces by using three-dimensional microfluidic systems. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[25]  A. Ito,et al.  Cell-patterning using poly (ethylene glycol)-modified magnetite nanoparticles. , 2009, Journal of biomedical materials research. Part A.

[26]  V. Yadavalli,et al.  Fabrication of poly(ethylene glycol) hydrogel microstructures using photolithography. , 2001, Langmuir : the ACS journal of surfaces and colloids.

[27]  A. Khademhosseini,et al.  Microscale technologies for tissue engineering and biology. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Robert Langer,et al.  Engineering systems for the generation of patterned co-cultures for controlling cell-cell interactions. , 2011, Biochimica et biophysica acta.

[29]  Y. Nahmias,et al.  Laser-guided direct writing for three-dimensional tissue engineering. , 2005, Biotechnology and bioengineering.

[30]  A. Khademhosseini,et al.  Directed assembly of cell-laden microgels for fabrication of 3D tissue constructs , 2008, Proceedings of the National Academy of Sciences.

[31]  Wook Park,et al.  Guided and fluidic self-assembly of microstructures using railed microfluidic channels. , 2008, Nature materials.

[32]  Shilpa Sivashankar,et al.  Enhanced cell viability and cell adhesion using low conductivity medium for negative dielectrophoretic cell patterning , 2010, Biotechnology journal.