Toward a clinical grade expansion of human mesenchymal stem cells: A complete serum-free, xeno-free culture system
Mark Weiss and David Fiorentini
Biological Industries Israel Beit Haemek Ltd.
Human mesenchymal stem cells (hMSCs) serve as new promising tool for regenerative medicine and cell therapy with the advantageous over other stem cells types, mainly due to their safety record, multipotent characteristic, the broad variety of their tissue sources and for being immuno-privileged. The presentation addressed the ability of a developed xeno-free culture medium as well as solutions for attachment, dissociation, and freezing to support long-term expansion of multipotent hMSC suitable for clinical applications.
Room 254 A
Cleaning up the mess: Isolation of pure germ layer derivatives
Sebastian Knoebel, Senior R&D Project Manager Stem Cells
Miltenyi Biotec GmbH
Standardized differentiation of pluripotent stem cells requires careful monitoring and selection of the desired cell progeny. Surface biomarkers help defining the phenotypic status and enable straightforward magnetic selection of distinct cellular subsets. Here, we discuss tools that facilitate efficient differentiation into each of the three developmental germ layers. The use of purified cell populations guarantees predictable and reproducible results and makes the stem cell workflow amenable to automation and clinical translation.
Room 257 A-B
Generating Parkinson’s Disease models using Footprint-free Reprogramming and a Novel Method for NSC Differentiation
Dr. Birgitt Schuele, MD, Assistant Professor, Clinical Molecular Geneticist, The Parkinson’s Institute, Sunnyvale CA
Life Technologies The ability to generate iPSC-derived models that recapitulate phenotypes associated with Parkinson’s Disease holds promise to accelerate understanding of the disease with a goal of finding a cure. By using non-integrating Sendai-virus (SeV) based CytoTune® reprogramming technologies, we have created patient-derived iPSCs of known genetic backgrounds, and used a novel method to create neural stem cells (NSCs) without going through the laborious process of EB formation followed by rosette isolation.
Gaining insights into tumor heterogeneity and cancer stem cell pathways by using high-throughput flow cytometry
Justin D. Lathia, PhD, Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic
Advanced cancers have a high degree of cellular heterogeneity and are often organized in a hierarchy with a self-renewing cancer stem cell (CSC) at the apex. A key issue for CSC studies has been the lack of consistent cell surface marker(s) for their prospective enrichment. To directly address this issue, we evaluated two advanced tumors (colorectal cancer and glioblastoma) using high throughput flow cytometry and have identified novel CSC pathways for future functional studies.
Room 258 A-C
Long-term call culture observation system and advanced image analysis technology
Dr. Lee Rubin, Harvard University
BioStation CT is an advanced, automated cell culture observation and documentation instrument. Ideally suited for live cell imaging, BioStation CT maintains a stable environment during microscopic image acquisition. This strategy and revolutionary new approach will lead to improvements in cell characterization methodologies and a better understanding of the reprogramming and differentiation process.
Novel, Animal-Free Fibronectin and Collagen-I ECM Mimetic Surfaces, Enabling The Defined Expansion of hMSCs and Other Primary and Adult Stem Cell Types
Deepa Saxena Ph.D.
As the utility of primary and adult stem cells progresses to clinical settings, the need for animal-free and scalable cell culture becomes a requirement. To enable clinical researchers achieve this, Corning has developed a line of ECM Mimetic Cultureware to support collagen-I or fibronectin dependent cell types. Here, we demonstrate expansion and functionality of hMSCs, keratinocytes and endothelial colony forming cells on ECM Mimetic surfaces. Scalable and compatible with multiple media, ECM Mimetic Cultureware is a ready-to-use option where animal-free and defined conditions are desirable.
Room 254 A
Integrated solutions for cell manufacturing and screening
Stefan Miltenyi, CEO and founder of Miltenyi Biotec
Miltenyi Biotec GmbH
Pluripotent stem cells hold great promise for cellular therapies and use in drug and toxicity screenings. Manufacturing of well-defined cell populations for therapeutic and screening purposes requires scalable workflows that allow closed-system or high-throughput processing. We have developed new platforms and instruments for single as well as multiple-parameter cell sorting and analysis. They help streamlining GMP-compliant cell manufacturing and integrate seamlessly into common liquid handling systems.
Room 257 A-B
Transcriptional and Epigenetic Dynamics During Specification of Human Pluripotent Stem Cells
Alex Meissner, PhD. Associate Professor of Stem Cell and Regenerative Biology, Harvard University, Broad Institute, Cambridge, MA
The genomic distribution of DNA methylation and diverse chromatin modifications within any cell reflects and partly determines its identity. Taking advantage of the orchestrated regulatory dynamics during the directed differentiation of human pluripotent cells we investigated the early events that contribute to the specification of all three germ layers. By carefully dissecting these initial transcriptional and epigenetic dynamics we may be able to better predict differentiation propensities and derive therapeutically relevant tissue types more efficient and safely.
Room 253 A-C
Flow cytometry analysis of cellular and functional phenotypes of disease in patient specific human IPSC derived cells
Paulina Ordonez, MD, Assistant Professor of Pediatrics, Department of Pediatric Gastroenterology, Hepatology and Nutrition, University of California San Diego, La Jolla, CA
Induced pluripotent stem cell (IPSC) technology is a powerful tool to model human disease in relevant cell populations. However, accurate detection of disease phenotypes will require analysis of sufficient patient lines to account for variation related to genetic background. In these regards, traditional methods of phenotypic characterization can be costly, time-consuming and not quantitative enough. Therefore, we have adapted several flow cytometry assays to rapidly screen for pathologic cellular and functional phenotypes in IPSC-derived cells.
Room 258 A-C
Two Unique Technologies For Neural Stem and Induced Pluripotent Cell Lines: The Synthemax® Self-Coating Synthetic Substrate and FloWell™ A Perfusion-Based Technology for Continuous Culturing.
Mark Rothenberg Ph.D.
This tutorial will introduce two unique technologies for stem cell research. The FloWell, allows for continuous perfusion of media and reagents from a Source Well to the Cell Well over a 3-day period in a 6-well format, removing the need for manual media exchanges. The second focuses on the Synthemax self-coating substrate. Both technologies are ideal for culturing stem cell lines including neural stem and induced pluripotent cell lines. The discussion will introduce both technologies and discuss relevant data.