Conference Registration, Materials Pick-Up, Morning Coffee and Tea

Moving Diagnosis Closer to the Patient: Smart Consumables For Novel Point-of-Care Solutions
Iris Prinz, Head of Sales and Business Development, STRATEC Consumables GmbH

Within the presentation we will focus on a Point-of-Care solution realized at STRATEC Consumables jointly with Femtonics. This Point-of-Care cartridge includes several challenges e.g. manufacturing of fluidics including opening mechanism for blisters by injection molding, content loading like dispensing and lyophilisation as well as several assembly steps in order to realize a diagnostic platform with the aim to detect respiratory diseases.

Mid-Morning Coffee and Tea Break and Networking in the Exhibit Hall

Networking Lunch in the Exhibit Hall -- Visit Exhibitors and View Posters -- Japanese Bento Box Lunch

Direct Ink Writing 3D Printing for Fabricating Ultra-deformable Microfluidic Electronic Devices
Michinao Hashimoto, Associate Professor, Singapore University of Technology and Design, Singapore

Three-dimensional (3D) printing has become the new standard to fabricate microfluidic devices. Stereolithography (SL) printing has been increasingly used to fabricate fluidic channels, but the fabricated devices are restricted by attainable channel dimensions and properties of photoresins. Integration of channels with electronic components is also difficult due to the mechanism of SL printing. In this work, we present direct ink writing (DIW) 3D printing as an alternative route to fabricating microchannels. DIW 3D printing allows extrusion-based patterning of silicone-based elastomeric materials (e.g., room-temperature-vulcanizing (RTV) silicone and addition-curing two-part silicone) on unique substrates such as elastomeric sheets integrated with electronic components (e.g., light-emitting diode chips and near field communication (NFC) tags) to enhance the functionality of the device. To highlight the advantage of DIW-based fabrication of microchannels, an ultra-deformable film-based microfluidic device with embedded liquid metals is fabricated to demonstrate the device conformability that addresses mechanical mismatch at the tissue-device interface. Overall, DIW 3D printing offers unique opportunities for the fabrication of microfluidic devices with advanced functions and will be beneficial for the development of flexible sensors and actuators.

PµSL, An Ultra-Precise 3D Printing, and its Application in Microfluidics
June Lu, Senior Engineer, BMF Asia

3D printing has been rapidly developed and well accepted in academic researches and different industries. However, obvious drawbacks of current 3D printing technologies still inhibit their further application and hence commercialization. Among them, resolution is one of the most mentioned concerns. Projection Micro Stereolithography technology (PµSL), with ground-breaking 2µm/10µm resolution, has been increasingly applied in both scientific researches and new product development. Compared to traditional methods such as injection molding and machining, PµSL substantially reduces the cost and shorten the lead time, by avoiding the mold fabrication or complicated programming. Compared to lithography, PµSL can avoid complex casting and strenuous assembly procedures, and able to build parts with more complex 3D structures as well (e.g., internal channels). Furthermore, PµSL can help to break the fetters of mind which inhibit the imagination of engineers. They can now reconsider the designs which were believed to be impossible with other methods. This report describes the principle and advantages of PµSL, then provides some case studies of its applications in microfluidics.

3D Bioprinting by “Watercolor”
Nelson Khoo, Chief Business Development & Sales Officer, Fluicell AB

Today, iPSC, stem cells, gene editing and bioprinting hold a scientist’s focus to improve research results, use better surrogate models to mimic in situ organs and, improve quality of life for patients. Many of the tools now available have made it possible to create such models, to replicate a tissue surrogate i.e., respiratory epithelium, liver models, pancreatic mini-organoids, etc. Fluicell AB is a 3D bioprinting company that provides the unique capability to deliver solutions and cells into a cell culture with high precision, without having diffusion or requiring a typical bioink. This gives pharma and biotech, as well as innovative research questions in world renown institutes, the opportunity to make and study heterogenic and biological relevant models, spheroids or mini-organoids. Our Biopixlar™ bioprinter is a novel platform that take advantage of a microfluidic based printhead to carefully deliver solutions and cells of interest according to requirements – offering you a “Lab in a Tip.” Representative examples will be shown to illustrate our take on bioprinting.

Mid-Afternoon Coffee and Tea Break and Networking in the Exhibit Hall

Healios’ eNK Cells: Research, Development and Manufacturing of an Innovative Treatment for Solid Tumors
Hardy TS Kagimoto, Founder, Chairman and CEO, HEALIOS K.K., Japan

HEALIOS K.K. (“Healios”) has developed engineered NK cells (“eNK” cells), a novel gene-edited NK cell therapy designed to possess excellent efficacy for solid tumors. Healios’ eNK cells are produced from a proprietary iPSC line, which has been genetically modified for enhanced anti-tumor activity. The eNK cell platform is uniquely differentiated from other approaches, having been engineered with five enhancements that are important for cancer killing activity. This comprehensive set of functional enhancements - and in particular unique migratory and immune cell recruitment features – position Healios’ eNK cells with the potential to penetrate solid tumors and, once there, initiate robust tumor killing activity. Preclinical studies have demonstrated superior antitumor activity of eNK cells in comparison to unmodified iPSC-derived and peripheral blood NK cells in vitro and in vivo. In addition, eNK cells have been generated in a high quality, clinical grade form and are currently manufactured in Healios’ in-house GMP facility at the Kobe Center for Medical Innovation utilizing a proprietary, automated, feeder-cell free 3D perfusion bioreactor system at an efficient scale of 100 billion cells per batch. Healios is continuing with its testing of eNK cells both in its laboratory facilities in Kobe as well as through joint research with several academic institutions. Healios has begun pre-clinical consultations with the PMDA and aims to begin clinical trials using eNK cells in 2024.

Oncolytic Vaccinia Vectors for Cancer Immunotherapy
Takafumi Nakamura, Associate Professor, Division of Molecular Medicine, Tottori University, Japan

Vaccinia virus, once widely used for smallpox vaccine, has been genetically engineered and used as an oncolytic virus for cancer virotherapy. The therapeutic index is successfully enhanced by stricter tumor-specific viral replication, stronger oncolytic potency, and optimized induction of antitumor immunity. Two viral proteins VGF and O1 contribute to viral replication and spread by activation of EGFR-dependent MAPK/ERK pathway. VGF- and O1-deleted vaccinia virus (MDRVV) inhibited pathogenic viral replication in normal cells without impairing therapeutic replication in tumor cells. Recently, fusogenic vaccinia virus (FUVAC) was discovered during plaque purification of the MDRVV. FUVAC has a nonsense mutation in the viral gene K2L, whereas other viral genomes are maintained in the MDRVV. Using a bilaterally tumor-bearing syngeneic mouse model, unilateral administration of FUVAC more efficiently enhanced CD8+ T cell infiltration and inhibited tumor growth in not only treated tumors but also untreated tumors than MDRVV. Interestingly, FUVAC reduced tumor-associated immune suppressive cells such as regulatory T cells, myeloid-derived suppressor cells and tumor-associated macrophages in the injected tumor. In accordance with the change for the better, the anticancer effects of FUVAC in both injected and non-injected tumors were completely suppressed by depletion of CD8+ T cells. Taken together, fusogenic vaccinia virus exerts more potent antitumor immune responses by modulating the tumor microenvironment than non-fusogenic vaccinia virus. Furthermore, the simultaneous expression of immune-modulating genes in FUVAC augmented the antitumor activity via inducing potent and durable antitumor immune responses following viral oncolysis. The combined properties have the potential to overcome oncolytic virus-resistant tumors which limit the clinical benefits due to tumor heterogeneity and the complexity of tumor microenvironment. Thus, FUVAC would be better therapeutic platform as a next-generation oncolytic vaccinia vector.

CRISPR/Cas9 Gene Editing of MIR155HG in Primary Human T Cells to Prevent Acute Graft-versus-Host Disease
Ramiro Garzon, Professor, The Ohio State University, United States of America

Acute graft-versus-host disease (aGVHD) is the primary cause of non-relapse mortality in patients receiving an allogeneic hematopoietic cell transplant. Previously we showed miR-155 is necessary for development of aGVHD. Thus, there is an unmet need to target miR-155 using different strategies. To overcome this problem, we propose to delete miR-155 host gene (HG) in primary T cells from human donors using CRISPR/Cas9 genome editing. We identified three pairs of guideRNAs targeting the promoter or exon 3 (Ex3) of MIR155HG, all of which exhibited expected deletion sizes confirmed by genomic PCR and significantly reduced expression of mature miR-155 transcript relative to non-targeting control guide. Digital droplet PCR and whole genome sequencing followed by CHURCHILL genome analysis was used to identify on and off target effects. To assess in vivo function, we performed xenogeneic aGVHD transplants using MIR155HG edited or control T cells injected into NSG mice. Recipients of MIR155HG deleted T cells displayed significantly lower aGVHD clinical scores (p<0.01) and significantly improved survival compared to mice receiving control T cells. In summary, our studies support the development of genome editing of MIR155HG in primary human T cells as a strategy to prevent aGVHD.

Ghost Cytometry for Diagnostic, Cell Therapy, and Pooled Screening, and Beyond
Sadao Ota, Associate Professor, University of Tokyo, Japan

Our groups focus on the development of high-speed, pooled, and multimodal measurement systems such as droplet microfluidics and flow cytometry by integrating hardware, biological, chemical, and information technologies to extract valuable information from cells or utilize functional cells for biomedical purposes. In this talk, I will first introduce a high-speed cell analysis and sorting technology developed based on an AI-driven approach for analyzing cellular image information without creating images (GC: ghost cytometry). We have realized GC-based cell sorters in both fluorescent and unlabeled (refractive index distribution information) versions and are developing practical applications for diagnostic, cell therapy, and screening applications. An increasing variety of machine learning methods of both supervised and unsupervised learning approaches adds flexibility to the analysis. As time permits, I would also like to introduce the concept of networked cell analysis platforms and realized technologies that we have recently developed, including a fast three-dimensional imaging flow cytometer.

Japanese Beer and Sake Reception in the Exhibit Hall -- Network with Colleagues and Engage with Exhibitors and Poster Presenters