Generation of Lymph-Node-on-a-Chip Device Utilizing 3D Cell Culture Bioprinting of Conduit Mimicking Towers

Abstract

In the modern immune oncology field, currently there are challenges for allowing for effective immunotherapeutic drug screening systems for immune cell populations in a biologically relevant design for process development and analytical development experimental systems. This challenge has yet to be effectively overcome without using animal models prior to moving towards clinical studies. Without successfully recapitulating in-vivo like settings ex-vivo, data collection and analysis for immunologic behaviors such as T-cell migration, B-cell germinal center formations and isotype class-switching are unable to be demonstrated, and thus limit the research and development procedures and results by preventing thorough characterization of immunotherapies prior to use in human patients within clinical trials. The problem causes increased costs prior to release of immune oncology drug pipelines, as well as extended time delays and use of resources for pharmaceutical, medical device and clinical companies within the industry and the field. One key solution that has been emphasized to resolve the challenge of 2D culture evaluation systems vs. biologically relevant settings of experimentation and data production such as that within in-vivo models has been to utilize bioprinting which allows for the generation of multi-dimensional structures within cell culture environments allowing for more accurate and efficient recapitulation of organic systems within the biology of the body. Historically bioprinting was inspired by developments in material transfer processes which operated at varying scales ranging from nano to macro. However, with most cellular housing/mimicking systems the scale tends to gravitate more towards the microscale and has proven to be a feasible target to be achieved when bioprinting structures and environments of this size (Guillotin, 2010). Over time and since its invention in 1983 the resolution and detail level that could be achieved by bioprinting has improved, and the characteristics and unique abilities of the technology have become far more validated and credible throughout the iterations and applications (Mironov, 2006). One of the unique properties of the bioprinting technology when it comes to creation of three-dimensional structures for culture, is its ability to print these three-dimensional structures in such a fashion that it allows for living cells to be printed within the structures themselves such that cells are bioprinted within void space but within the volume of the device (Kolesky, 2014). This technology has allowed for better recapitulations of varying biological structures required to form significant organs within the body, one such being the lymph node through generating representative structures found within the organ that generate required passive and active signaling pathway triggers for natural cell behavior to take place. By having such recapitulation, mechanical, chemical, and systemically impactful cues are able to be generated through utilization of tonic signaling and the formation of concentration gradients. This approach will allow for cells to interact, react, and produce chemokines, cytokines, and mobility more representative to their standard state in which they do so within the body in-vivo, and do so ex-vivo posing advantageous properties as opposed to traditional two-dimensional in-vitro systems. With a three-dimensional system in place, compound and immunologic screening will be able to better characterize the effectiveness and efficiency of immunotherapies on cell populations within the body that are relevant for disease areas such as oncology and cancer treatment and prevention, autoimmune disease, and sickle cell disease. This demonstration will give more physiologically relevant data prior to, or in place of animal models, and allow for a better understanding of the impact on the immune system within human patients, and thus a clearer understanding of which immunotherapies screened, developed, and delivered will be the best product and most helpful in curing the disease in focus during clinical trials both for short-term and long-term evaluation. By having a system in place that allows for effective drug screening of immune oncology compounds and therapies, both a broad array of immune cells as well as cells of cancers, autoimmune disease, sickle cell disease, and other can be thoroughly evaluated and characterized through data collection and analysis in lieu of animal models so as to provide for quicker turnaround times for patients as well as more thorough evaluations of high-efficacy candidates. Future work of the technology’s broader implications can in turn be used for high-demand immunotherapies such as evaluation of combination therapies, as well as CAR-T screenings, characterization, and evaluation helping to generate product sets that target disease areas with high unmet need such as cancer in the solid tumor space for example. To establish the foundation of this technology, a bioprinted 3D cell culture will first be optimized, and then a series of co-cultures of relevant immune populations will be conducted to generate indicative data results and analysis demonstrating effectiveness.