Dear All, Below please find the second issue of the newsletter of the 3D Cell Culture Core (3D3C) Facility of the Birck Nanotechnology Center. The newsletter is also available online (https://nanohub.org/groups/3d3cfacility/news). There are four sections in the newsletter: 3D at Purdue – this section highlights 3D cell culture-based research activity at Purdue 3D in discussion – this section presents the current work on a specific 3D cell culture model or techique 3D in publication – this section brings a collection of recent publications on 3D cell culture 3D in meeting – this section includes a list of upcoming meetings related to 3D cell culture The newsletter will be available every two months. If you do not wish to receive the 3D3C newsletter in the future, please reply “cancel” to unsubscribe. Please contact me if you have questions. Yours Sincerely, Tim Kwok Facility Manager 3D Cell Culture Core (3D3C) Facility Birck Nanotechnology Center Purdue University ________________________________ [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/3e8e050f-67fb...] Volume 2, February 2016 [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/10f15e72-d4d6...] 3D at Purdue 3D in Discussion 3D in Publications 3D in Meetings 3D at Purdue In the Ziaie Biomedical Microdevices Laboratory (ZBML), we combine the design concepts of MEMS/BioMEMS technology with modern rapid prototyping equipment to develop novel devices and microsystems that address important clinical problems. We use equipment such as laser engravers, cutter-plotters, and 3D printers to process and repurpose a multitude of materials which are exotic to traditional MEMS fabrication but invaluable for medical applications. Such materials include ultra-soft elastomers (for conforming to tissue/organs), a broad range of functional polymers with unique surface properties (for automatic response to changing microenvironments), as well as paper and fabrics (for creating inexpensive, wearable, and disposable diagnostics/therapeutics). 3D printing is particularly advantageous for creating structures with complex morphologies which mimic the intricate micro-architectures found in nature (e.g., micro-scales or micro flanges in insects and plants). One recent application of 3D design in our lab is a practical fabrication technique for creating polymeric microneedles of complex geometries for drug/vaccine delivery applications, Figure 1 [1]. This technique allows the creating of more sophisticated, slanted, or out-of-plane designs that allow for easier needle insertion using smaller shear force. We achieve this by coupling 3D printing technology with an isotropic shrinkage technique, which effectively enhances the current resolution limits of 3D printing by at least five fold. The resulting needles are sufficiently sharp to penetrate porcine skin and deliver loaded/embedded chemicals. [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/33a1d70a-3f99...] Figure 1: Photographs of polymeric microneedles. (a) and (c) show two 3D printed molds of different sizes; (b) and (d) show shrunk replicas of each mold after two gel-shrinking iterations. Tip radius of curvature (boxed in yellow) is (a) r = 212 µm; (b) r = 14.7 µm; (c) r = 56 µm; (d) r = 9.6 µm. (e) Final dissolvable polymeric needles. (f) Alternate complex 3D mold. We are also investigating the use of 3D architectures for cancer research in collaboration with Prof. Lelièvre. By combining the diverse but complementary expertise of both labs, we are engineering device platforms for culturing cancer cells in a 3D environment which closely mimics the human physiology in order to create improved tumor models for evaluating anti-cancer drugs. One example of this is the development of laser-machined hemi-microchannels as models of mammalian breast cancer cells, Figure 2 [2]. By culturing cells along the curved walls of a hemichannels, their physiology is able to more accurately mimic tumor development within the epithelial environment of mammary ducts, providing a framework for the design and test of anticancer therapies. [https://gallery.mailchimp.com/b7d6d1ffee56a866e499104cf/images/2d851ff5-a00d...] Figure 2: Monolayer of cells on a smooth semicircular acrylic channel. SEM images (left) show the appearance of each type of hemichannel before seeding the cells. Nuclei are stained with DAPI (blue). Scale bars, 10 μm. We look forward to accelerating the pace of high-impact medical research by using such rapid prototyping fabrication technologies along with fruitful inter-disciplinary collaborations at Purdue. Babak Ziaie, PhD Professor, Electrical and Computer Engineering [1] M. Ochoa, J. Zhou, R. Rahimi, V. Badwaik, D. Thompson, and B. Ziaie, “Rapid 3D-print-and-shrink fabrication of biodegradable microneedles with complex geometries,” in Proc. Transducers - 2015 (Anchorage, AK), 2015. [2] P.-A. Vidi, T. Maleki, M. Ochoa, L. Wang, S. M. Clark, J. F. Leary, and S. A. Lelièvre, “Disease-on-a-Chip: Mimicry of Tumor Growth in Mammary Ducts,” Lab Chip, 2013. 3D in Discussion Research Paper: Wang JD, Khafagy ES, Khanafer K, Takayama S, ElSayed ME. Organization of Endothelial Cells, Pericytes, and Astrocytes into a 3D Microfluidic in Vitro Model of the Blood-Brain Barrier. Mol Pharm. Epub ahead of print DOI: 10.1021/acs.molpharmaceut.5b00805 The blood–brain barrier (BBB) consists of endothelial cells lining the brain capillaries system and surrounding astrocytic glial membranes. The barrier function is dependent on the tight junctions between the endothelial cells and the specific transport proteins in the cell systems. The BBB lets some substances, such as water, oxygen, carbon dioxide, and general anesthetics, pass into the brain. It also keeps out bacteria and other substances, such as many anticancer drugs. The BBB is the bottleneck in brain drug development and is the single most important factor limiting the growth of neurotherapeutics. The BBB can be measured by in vivo, in vitro, and in silico system (Bickel U, J Am Soc Exp NeuroTherapeutics, 2005, 15 - 26). An effective in vitro model for BBB will allow their use for noninvasive, rapid, economic, and reproducible screening of the BBB permeability of new drug candidates. However, the current in vitro BBB models fail to replicate the organization and restrictive behavior observed in vivo due to poor integration of the endothelial cells with supporting cells (pericytes and astrocytes) following the correct anatomical organization observed in vivo. The manuscript described an in vitro BBB model by co-culture of mouse brain microvascular endothelial cells, pericytes, with astrocytes in layered microfluidic channels forming three-dimensional models of the BBB. The model exhibits different degrees of “restrictiveness” indicated by their high transendothelial electrical resistance (TEER) values, low permeability of [14C]-mannitol and [14C]-urea, and functional expression of the P-glycoprotein efflux pump. Based on the similarity in mannitol permeability across cell monolayers in the BBB model with the permeability reported for the BBB in vivo and the high functional expression of the P-glycoprotein efflux pump, the tri-culture BBB model described in this report represents a 3D in vitro model of the BBB that closely mimics its restrictive nature observed in vivo. The work described in the manuscript is a typical case for 3D tissue model development, the crossover of cell biology and engineering technology. 3D in Publications Recent publications on 3D culture (please click to access the list on our web page https://nanohub.org/groups/3d3cfacility): Review<https://nanohub.org/groups/3d3cfacility/news#reviews> The research papers and reviews are arranged in the following category: Scaffold free/Scaffold Organ/Tissue/Cell Others Spheroids<https://nanohub.org/groups/3d3cfacility/news#spheroids> Organoid<https://nanohub.org/groups/3d3cfacility/news#organoid> Scaffold<https://nanohub.org/groups/3d3cfacility/news#scaffold> Hydrogel<https://nanohub.org/groups/3d3cfacility/news#hydrogel> Matrix<https://nanohub.org/groups/3d3cfacility/news#matrix> Microfluidics<https://nanohub.org/groups/3d3cfacility/news#microfluidic> Microfabrication<https://nanohub.org/groups/3d3cfacility/news#microfabrication> Adipocyte<https://nanohub.org/groups/3d3cfacility/news#adipocyte> Bladder<https://nanohub.org/groups/3d3cfacility/news#bladder> Bone<https://nanohub.org/groups/3d3cfacility/news#bone> Bone Marrow<https://nanohub.org/groups/3d3cfacility/news#bonemarrow> Breast<https://nanohub.org/groups/3d3cfacility/news#breast> Colon<https://nanohub.org/groups/3d3cfacility/news#colon> Heart<https://nanohub.org/groups/3d3cfacility/news#heart> Liver<https://nanohub.org/groups/3d3cfacility/news#liver> Lung<https://nanohub.org/groups/3d3cfacility/news#lung> Muscle<https://nanohub.org/groups/3d3cfacility/news#muscle> Nerve<https://nanohub.org/groups/3d3cfacility/news#nerve> Prostate<https://nanohub.org/groups/3d3cfacility/news#prostate> Endothelial cells<https://nanohub.org/groups/3d3cfacility/news#endothelialcells> Fibroblast<https://nanohub.org/groups/3d3cfacility/news#fibroblast> Stem Cells<https://nanohub.org/groups/3d3cfacility/news#stemcells> Stromal Cells<https://nanohub.org/groups/3d3cfacility/news#stromalcells> Plant Cells<https://nanohub.org/groups/3d3cfacility/news#plantcells> Cancer/Tumor<https://nanohub.org/groups/3d3cfacility/news#cancer> Screening<https://nanohub.org/groups/3d3cfacility/news#screening> 3D bioprinting<https://nanohub.org/groups/3d3cfacility/news#3dbiprinting> Imaging<https://nanohub.org/groups/3d3cfacility/news#imagining> Quantitation<https://nanohub.org/groups/3d3cfacility/news#quantitation> 3D in Meetings EMBO|EMBL Symposium: Tumour Microenvironment and Signalling Conference 3rd to 6th April 2016 EMBL Advanced Training Centre, EMBL Meyerhofstrabe 1, Heidelberg 69117, Germany Website: http://atnd.it/39532-0 Contact person: Course and Conference Office This symposium brings together researchers from complementing fields to enhance our understanding of the communication between cancer cells and their microenvironment. Organized by: European Molecular Biology Laboratory, EMBL Heidelberg World Stem Cells and Regenerative Medicine Congress 2016 Conference 18th to 20th May 2016 London, United Kingdom Website: http://atnd.it/36213-0 Contact person: Katy Scrivener Europe’s market place for investment, commercial opportunities and collaboration. Focusing on commercialising cell therapies, stem cells for drug discovery, tissue engineering and organ regeneration. Time: £50 - £3070. Organized by: Terrapinn 2016 International Forum on Medical Physics, Biomedical Engineering and Biotechnology Conference 14th to 15th July 2016 Osaka, Japan Website: http://www.astfi.org/IFMBB2016/index.html Contact person: Christopher Zhu The conference aims to foster and conduct collaborative interdisciplinary research in state-of-the-art methodologies and technologies within Medical Physics, Biomedical Engineering and Biotechnology. Organized by: Asia Science and Technology Forum Institute EMBO|EMBL Symposium: Organoids: Modelling Organ Development and Disease Conference 12th to 15th October 2016 Heidelberg, Germany Website: http://atnd.it/39583-0 Contact person: Course and Conference Office This conference will bring together the leading researchers in organoid field to establish a new research community and reveal parallels between various tissue models. Organized by: European Molecular Biology Laboratory, EMBL Heidelberg