2014 Archived Content
Dinner Short Courses*
SUNDAY EVENING, NOVEMBER 16
(SC1) Introduction to High-Content Phenotypic Screening
The ever-increasing demand for improved productivity in research through the generation of robust analysis outputs has driven both the development and deployment of automated high-content analysis (HCA) and phenotypic cell-based approaches to drug discovery. In contrast to the more traditional cellular analysis and target-based approaches, here the researcher is able to evaluate the efficacy of potential therapeutics by monitoring the physiological state of cells through the simultaneous analysis of multiple cellular parameters in the context of an intact biological system. This course will cover the key features of HCS/A technologies and the best approaches to using these technologies for phenotypic cell-based screening.
Anthony M. Davies, Ph.D., Center Director, Translational Cell Imaging Queensland (TCIQ), Institute of Health Biomedical Innovation, Queensland University of Technology
Who should attend?
This course has been developed to introduce and facilitate scientists who are either moving into the field or who are interested in further developing new phenotypic discovery applications and tools for use with these technologies.
(i) An introduction to HCA technologies
(ii) Advanced cell-based models for use with HCA
(iii) Worked examples of the phenotypic screening approach and future directions
(iv) Group discussion and Q&A
- Develop a familiarity of the basics of HCS/A technologies
- Gain an understanding of the capabilities of this technology
- Learn of the latest developments in cell-based models for use in this field
- Get a better understanding of the key principles of assay design and development for phenotypic screening
(SC2) Exploring 3D Printing, Bioinks and Scaffolds
The promise of 3D bioprinting to create human tissues layer by layer is immense, ranging from basic biological research to drug development and testing, and ultimately to replacement organs. However, organ and tissue structures vary in complexity, and printing with living cells to create tissues is much more complicated than printing 3D objects in plastic.
Topics to be covered:
- 3D Printer Platforms: Inkjet vs. Pressurized Printing
- 3D Modeling (CAT Scans, Laser Scans and CAD)
- Scaffold Selection
- Cell Source Selection
This dinner course is designed for biological researchers who are interested in learning more about 3D printing and applying it to building a living tissue or organ of their choice.
6:15 Future Medical Applications in 3D Printing: Clinical Benefits, Regulatory Issues and Manufacturing Challenges
Michael Drues, Ph.D., President, Vascular Sciences
What are the future medical applications in 3D printing? Can we print actual medical devices and permanent implants? Can we print drugs and living tissue? What are technical and regulatory challenges, and what’s holding us back?
6:50 Dinner Break
7:15 2D Printing to 3D Tissue Fabrication via Multilayer Stacking of Biopapers
Russell Kirk Pirlo, Ph.D., Research Biologist, Chemistry, U.S. Naval Research Laboratory
There are several two-dimensional cell and biofactor printing technologies available today. Stacking and/or multilayer printing allows these technologies to be used to create three-dimensional tissue constructs. I discuss the general challenges and considerations in selecting and designing cell-ink and scaffold materials as well a method for inter-layer registration of individually addressed substrates. Detailed reference to our own Biological Laser Printing and Biopaper technologies will be used as an illustrative case study.
7:50 Multimaterial 3D Bioprinting
David Kolesky, Research Scientist, Jennifer Lewis Laboratory, School of Engineering and Applied Sciences and Wyss Institute for Biologically Inspired Engineering, Harvard University
To vastly expand the application of 3D bioprinted tissue constructs, one must be able to integrate cells, structural and vascular components concurrently and with precision. Using a custom-designed bioprinter, we are creating 3D multimaterial, cell-laden architectures with embedded vascular networks. I will describe the printing platform and our enabling ink designs as well as characterization of printed 3D cells and microstructures.
8:25 The Organovo 3D Bioprinting Platform: Changing the Shape of Medical Research and Practice
Deborah G. Nguyen, Ph.D., Director, R&D, Tissue Applications, Organovo, Inc.
The high failure rate among clinical-stage therapies underscores the need for improved in vitro models. A common platform to produce both preclinical and therapeutic tissues would be immensely attractive. Organovo’s 3D Bioprinting platform generates microscale tissues that mimic human tissue architecture and function without exogenous biomaterial scaffolds. We will explore how medical research and practice challenges are being addressed by this novel technology platform.
9:00 Close of Short Course
MONDAY EVENING, NOVEMBER 17
(SC3) Stem Cell Models for Drug Discovery
Induced Pluripotent Stem Cell-Based Disease Models in Drug Discovery
Anne G. Bang, Ph.D., Director, Cell Biology, Prebys Center, Sanford-Burnham Medical Research Institute
Phenotypic screens of human neurons from patient-specific induced pluripotent stem cells (iPSC) can be used as discovery tools to generate new hypotheses regarding mechanisms underlying neurological disease and identify treatments based on biology rather than symptoms. An important step is to establish the technology platforms and reproducibility necessary to utilize iPSC in high-throughput drug screening. Towards this goal, we have been working to develop a standardized battery of assays against which iPSC-derived neurons can be screened for specific phenotypes. We have performed a high-content screen at the scale of 26,000 wells for chemical modulators of neurite growth using hiPSC-derived neurons, and have developed a miniaturized multiplexed high-content assay to assess neuronal ROS and mitochondrial potential. In addition, we have conducted a 5,000 well screen using Alzheimer patient-specific iPSC-derived neurons. Finally, we are developing assays to analyze neuronal function using multi-electrode arrays on multi-well plates. We will discuss our screening results and development of iPSC-based models for testing of drugs on disease-relevant cell types.
Human Intestinal Epithelium Derived from Induced Pluripotent Stem Cells
Pamela J. Hornby, Ph.D., Senior Scientific Director and Research Fellow, Cardiovascular and Metabolic Disease, Translational Models, Janssen Pharmaceutical Companies of Johnson & Johnson
Intestinal organogenesis methods (McCracken, et al., 2011; Rezania, et al., 2012) were optimized to differentiate five retrovirally and non-virally reprogrammed human induced pluripotent stem cells (iPSCs) lines. These 3D organoids expressed markers of epithelial lineages to varying degrees. One retrovirally reprogrammed cell line was adapted to culture in transwells and monolayers with high TransEpithelial Electrical Resistance and low FITC-Dextran permeability. Adoption of standard protocols to improve robustness of organoid formation may make iPSC-derived intestinal-like cells a useful tool for understanding healthy and diseased epithelial mechanisms.
Application of Patient Neuronal Cells Differentiated from iPS Cells as Disease Models for Phenotypic Screening
Wei Zheng, Ph.D., Group Leader, National Center for Advancing Translational Sciences, National Institutes of Health
Phenotypic screening has reemerged as an effective approach for compound screening to identify lead compounds for drug development. Recent advancement in iPS cell technology has enabled quick generation of induced pluripotent stem (iPS) cells from patient skin cells that can be further differentiated to mature cells such as neuronal cells as a disease model system for compound screening. This phenotypic disease model system is particularly useful for these diseases of which the disease pathophysiology is unclear and drug targets are not available. We have generated several iPS cell lines from patient cells with lysosomal storage diseases including Niemann Pick disease Type C, Wolman Disease and Mucopolysaccharidosis type-I. The neuronal cells differentiated from these patient iPS cells exhibit characteristic features of lipid/macromolecule accumulation and enlarged lysosomes. The results from compound screening and drug efficacy tests using these differentiated disease cells will be presented and discussed.
(SC4) Engineering Microfluidic Cell Culture Chips
Microfluidic technology holds great promise for the creation of advanced cell culture models. Engineering a microfluidic cell culture chip to emulate the dynamic physiology of a tissue’s microenvironment is paramount for primary cell culture and co-culture. As the availability of functional human cell types for in vitro culture increases, a microfluidic cell culture chip platform’s potential to produce an in vitro system capable of accurately reproducing acute and chronic human responses to drug and pathological challenges in real time will also increase.
Topics to be covered:
- Cell Source Selection
- Media Requirements
- Channel, Chamber and Valve Design
- Characterization of Microfluidic Systems
- Integration of Analytics
- Achieving High Throughput
This dinner course is designed for researchers interested in delving into the world of microfluidic cell culture to determine its advantages over their traditional cell culture.
6:30 pm Welcome
6:50 Dinner Break
7:20 The Basics of Integrating Cells with Microfluidic Devices for Long-Term Cell Survival and Function in Organ-on-a-Chip Devices
James J. Hickman, Ph.D., Professor, NanoScience Technology, Chemistry, Biomolecular Science and Electrical Engineering, University of Central Florida
Defined systems are the best option for integrating cells and tissue with microfluidic devices in controlled, reproducible assays. A defined system is composed of a known surface chemistry, serum-free medium and knowledge of cell type, differentiation state and preparation history. Examples of defined systems for cardiac, neuronal and liver as well as methods of characterization will be presented.
8:10 Refreshment Break
8:20 Fabricating a Liver Tissue Model in Microfluidic Platforms
Rohit Jindal, Ph.D., Instructor, Surgery, Center for Engineering in Medicine, Massachusetts General Hospital
I will discuss various issues that are involved in creating a liver tissue model in microfluidic devices. I will also describe our recent results pertaining to a mechanism through which flow stabilizes primary hepatocyte cultures in microfluidic systems.
9:10 Interactive Q&A with Instructors and Participants
9:30 Close of Short Course
TUESDAY EVENING, NOVEMBER 18
(SC5) Expert ThinkTank: How to Meet the Need for Physiologically-Relevant Assays?
It used to be adequate to build target-specific and robust assays to drive lead optimization. These assays were relatively inexpensive and reliable and could be counted on to provide chemists with usable results. However, with time, it has become apparent that it is not enough to be robust and target specific. To build therapies for patients, we need to have assays that are more predictive of patient outcome. The current buzz words are "physiologically-relevant assays." This session will explore the need for physiologically-relevant assays and explore the ways that we can achieve this endpoint.
Lisa Minor, Ph.D., President, In Vitro Strategies, LLC
- Beverley Isherwood, Ph.D., Team Leader, AstraZeneca R&D
- Michael Jackson, Ph.D., Senior Vice President, Drug Discovery and Development, Conrad Prebys Center for Chemical Genomics, Sanford-Burnham Medical Research Institute
- Jean-Louis Klein, Ph.D., Principal Scientist, Target and Pathway Validation, Platform Technology and Science, GlaxoSmithKline
- Caroline Shamu, Ph.D., Director, ICCB-Longwood Screening Facility and Assistant Professor, Harvard Medical School
- D. Lansing Taylor, Ph.D., Director, University of Pittsburgh Drug Discovery Institute and Allegheny Foundation; Professor, Computational and Systems Biology, University of Pittsburgh
- Scott S. Verbridge, Ph.D., Assistant Professor, School of Biomedical Engineering and Sciences, Virginia Tech – Wake Forest University
Discussion topics include:
- What is the driver for more physiologically-relevant assays?
- What is the current state of the art?
- Advantages and shortcomings of the current technologies
- What is needed to build more physiologically-relevant assays? What are the requirements for cells, assay systems and budget?
- What are the compromises that can be allowed when building tools?
- What would be the ideal outcome and timelines?
*Separate registration required