Interviews

Interview with Rebecca A. Lead, Postdoc in CONNECT Nervous-system-on-Chip

Dr. Rebecca A Lea has been a postdoctoral research associate in the Tsakiridis Lab, studying the biology of the vagal neural crest and its downstream derivative, the enteric nervous system (ENS). Her passion is to understand the underlying biology of neurodegenerative disease, in the hope that this will pave the way to future treatments and increased quality of life for patients. She received an MBiochem in Molecular and Cellular Biochemistry from the University of Oxford, UK in 2016 and went on to complete a PhD in Stem Cell and Developmental Biology at the Francis Crick Institute and University College London, UK in 2021. Following on from her PhD work, in which she investigated the basic biology of human pluripotent stem cells (hPSCs), she moved to Sheffield to join the CONNECT project under Dr. Anestis Tsakiridis, to apply her expertise in hPSC biology to cutting-edge neurodegenerative disease modelling.

You are moving on to a new position, what was the objective of your postdoctoral studies in CONNECT?

During my time on the CONNECT project, I worked to investigate the feasibility of using an integrated ENS-intestinal organoid model as a starting material for the on-chip system. Our hypothesis was that by culturing ENS components in a way that more closely mimics their in vivo environment, we could improve the quality and functionality of the desired enteric neurons and glia. Unfortunately, maintaining the co-cultures in the long-term proved unfeasible, but I still gained invaluable experience in 3D organoid culture, and we now know that monolayer ENS cultures will be best-suited to the CONNECT chip systems.

What do you think (from the viewpoint of your respective scientific discipline) is the biggest challenge in combining chip technology with neurobiology?

I think one of the major challenges in all disease modelling applications is how to know whether, or how well, our model faithfully represents the physiological environment, i.e., in the human body. With chip technology, we have the advantage of being able to precisely modify various parameters to tweak the ways in which our models form and grow. However, this is still limited by our relative lack of knowledge about the native microenvironment of the desired cells, e.g., in the brain. Testing a variety of chip configurations, alongside different raw materials and substrates, for each component of the nervous-system-on-chip model is likely to be a time-consuming process, but may be vital for its successful application.

You have been talking part in CONNECT – Collegial Start-up contest in 2023, if you were now meeting an investor in an elevator, what would be your 15 seconds Message Map to pitch the importance of putting the next level in vitro model systems for the nervous system firmly on the map?

Interview with Yi-Ning Kang, PhD student in CONNECT Nervous-system-on-Chip

Yi-Ning is a PhD student in the group of Prof. Pieter Van den Berghe at KU Leuven (Katholieke Universiteit Leuven, Belgium). She had her two-year master training on live cell imaging in National Yang-Ming University, Taiwan. Her master thesis had won the first place of Outstanding Thesis Award among nearly 600 graduates. In 2019, she decided to keep following her interests on advanced imaging techniques with the guidance from Prof. Pieter Vanden Berghe in Leuven. For her current PhD research, she is taking advantage of the CONNECT Nervous-system-on-Chip design to answer substantial questions about the development of the enteric innervations.

What is the objective of your project? What outcomes do you expect after completing your second year of your PhD studies?

I will use advanced live cell imaging techniques to characterize and validate the physiology of the artificial enteric nervous system. We aim to ensure that the functionality of hPSC-derived enteric nervous system in terms of synaptic transmission and neuronal activity is comparable to the in-situ one. Hopefully after completing the second year of my PhD research, I would be able to see the transport of mutant alpha-synuclein (toxic aggregates) between different compartments on the CONNECT chip, in which different nerve networks have been mimicked.

What comes to your mind when you hear brain modelling and why do you think it makes a difference to apply chip technology to unravel the mechanism of Parkinson’s disease and find new treatment methods?

I can imagine how hard it would be to model human brains. Our brain is such a complicated organ that it controls most of human basic functions. However, if we manage to develop an organ-on-chip design to mimic certain relevant aspect of the brain, or other nerve networks, we would be able to answer a lot more critical questions on the mechanism of neurodegenerative diseases. Not to mention that most of the animal studies are time-consuming and do not recapitulate the human disease very well. The CONNECT chip would provide a fast and easy platform for us to use human cells and to test different potential drugs for treatments.

How is the collaboration within this project going? How do you think the radial microtunnel chip made by Rahman can help provide answers to specific research question in your field?

Even though everything becomes harder due to corona, luckily, I have supportive partners within the CONNECT project. They are always kind and thoughtful to solve my questions. Moreover, they trust my expertise on optical recordings and wait for my results with patience. The radial microtunnel chip made by Rahman well fits the idea of one nerve network connects to many others. Recently, with that chip and the review that I published (see review), in which I summarize various molecules that may play crucial role during the development of the enteric innervations, I can use one compartment as a control, others with specific treatments, in order to see what molecules indeed attract the development of enteric innervations.

Are you planning to attend a conference presenting your research results soon, even this may be online? If yes, please tell us a bit more about the type of conference and who the main attendees are? Are there also participants from industry, representatives of patient groups or medical specialists?

I am not planning to attend any conference to give a presentation of my research results at the moment. Since the outbreak of pandemic, my research has been strongly affected and has progressed dramatically slow, which didn’t fit my expectation of a one-year research findings. Therefore, I rather do more practical experiments and attend conferences or summer schools online to advance my knowledge and skills, but not to present my research results. However, I am confident that soon experimental work will be more normal and as soon as I discover something exciting, I am more than happy to show everyone.

Interview with Rahman Sabahi Kaviani PhD student in CONNECT Nervous-system-on-Chip

Rahman has obtained his B.Sc. degree in Mechanical Engineering from Sharif University of Technology, Iran, where he graduated in the top 5% of his class. Later, he received his M.Sc. degree in Mechanical Engineering from the University of Michigan, Ann Arbor, MI, USA, where he was nominated for the prestigious Rackham Fellowship award. He then continued his research in BioMEMS laboratory in the University of Michigan and Lurie Nanofabrication Facility, where he gained further knowledge and hands-on experiences in design and fabrication of microfluidic devices and microsystems. Prior to joining Eindhoven University of Technology for his PhD studies, he gained further research, industrial and teaching experiences in General Motors company, Iran’s National Elite Foundation and Sharif University of Technology.

What is your specific technical contribution to the objective of CONNECT project, so far? What outcomes do you expect after completing your second year of your PhD studies?

In CONNECT, we are developing an in-vivo like microenvironment that can bring together cells and organoids resembling different parts of the human nervous systems. I recently proposed microtunnel devices to provide such capabilities. These chips allow us to direct axonal growth and neuronal network formation in a compartmentalized fashion. For example, my fellow PhD student in the CONNECT project at KU Leuven, Yi-Ning, currently investigates these chips for the growth of enteric neurons. Moreover, I contributed to the ongoing research on microsieves in our group by advancing pattern fidelity of 3D micropores using replica molding and excimer laser micromachining (see article). These devices aim to extend our chip technology toolbox to enhance differentiation processes of nervous systems’ stem cells. I can now robustly fabricate the microsieves to be ready for further culture tests at our partners lab. By completing the second year of my research project, I expect to demonstrate an early prototype of the microfluidic CONNECT chip platform also implementing the concept of surface modification in such devices utilizing, for example, nanoimprinting of nanogrooves. I also aim to apply for my first patent application on such an integrated devices functionality by the proposed microfabrication methods in my PhD studies.

What comes to your mind when you hear about 3D organoids and novel technical routes to model the brain? And, why do you think it makes a difference to apply microtunnel devices integrated into microfluidic brain-on-chips in the study of the nervous system?

When I hear about 3D organoids, I think of an in-vitro 3D object which is the aggregation of cells derived from stem cells and can express the functionality of specific tissue or organ. The novel technical routes to model the brain, not only needs to benefit from novel biological protocols for preparing such specific neuronal cells or 3D organoids from stem cells, but also needs to deploy advanced technical solutions to produce a more realistic (physiological) environment. Microtunnel devices are offering special features, like compartmentalization, in the study of the nervous systems, which make them desirable structures. They can connect cells and organoids with different identities and reduce nutrients flow by the limited diffusion from one compartment to the other. Next, these devices also provide a region for a clear optical observation of molecular transport along the connecting axons.

How is the collaboration within this project going? Have your chips been tested already? Why is this exciting?

As mentioned in my answer above, CONNECT is a highly interdisciplinary project. The collaboration between consortium partners with different research areas is an essential part of the work. These interactions are very important when an element in the platform is designed and fabricated and needs to be tested by our biological expert partners for culturing specific cells and organoids. The microtunnel device has been previously tested for SH-SY5Y cell culture in our group (see article), but to test the performance with relevant neuronal cell types, they have been dispatched to our partners in University of Luxembourg, KU Leuven an Erasmus MC. The preliminary results of our partners’ experiments are already very exciting and confirm that the microtunnel device can be beneficial to the further integration of the CONNECT chip platform. We also have our regular technical meetings with our partner in Aalto University for investigating the possible solutions for advanced integration of electrochemical and electrical sensing electrodes into these chips as a next step in our developments.

I have submitted an abstract of my recent progress to the 64^th international conference on Electron, Ion and Photon Beam Technology and Nanofabrication (EIPBN). I hope my work gets accepted and I am eager to share my outcomes within this long-standing community of micro- and nanofabrication specialists, bringing together professionals from academia, industry, and governmental research institutes from all around the world. This conference is recognized as the foremost international meeting dedicated to lithographic science and technology. Although this year’s conference is virtual, it will still be a great opportunity for me to interact with delegates and commercial exhibitors dedicated to my field of research and discuss future trends.

In your own words, how do you think does CONNECT make a difference to society?

CONNECT will provide a more realistic brain model on a chip by connecting different neuronal cells and organoid. The fact that all of the biological components will be derived from human stem cells, not only makes the model representing a human brain, but also eliminates the need for the morally debatable animal use for pre-clinical studies. The platform is intended to model Parkinson’s disease first, but successful outcomes could also be implemented for other neurodegenerative diseases. Therefore, the successful completion of CONNECT will aid toward better understanding of the brain and brain diseases and the development of novel drugs and medicines.

Is there something else you would like to share?

CONNECT has a very ambitious goal to achieve, however, along the way, there are very interesting midway goals to gain. Specifically, in my line of research, I have investigated and produced different components, and will design and come up with other parts needed for the final platform. This learning route in addition to our frequent interactions with our partners and learning from their perspectives, have been very educational and exciting. Although the COVID-19 pandemic has hindered and delayed some experimental works, thanks to the openness and dedication of all PhD’s, Post docs and PI’s involved in the project, the consortium keeps up progress.

Interview with Samuel Rantataro, PhD student in CONNECT Nervous-system-on-Chip

Samuel Rantataro is a PhD student at Aalto University, Helsinki (Finland) in the Department of Electrical Engineering and Automation, who works together with professor Tomi Laurila and Sami Franssila towards his PhD on the integration of highly sensitive electrodes for detecting dopamine and other neurotransmitters in physiologically relevant environments. He received his M.Sc. in Materials Science from Aalto University in 2019, in which he focused on functional materials and micro-/nanotechnologies. Prior to joining CONNECT, he has successfully worked on various microfabrication projects related to MEMS, microfluidics, and process design. He became interested in combining his earlier knowledge with neuroscientific research, resulting into an awarded Master’s (link) in studying the effect of substrate stiffness on neural cell mechanics. Now he focuses on electrochemistry of neurotransmitters, while also on integrating electrochemical electrodes into Nervous-system-on-Chip technologies.

What is the objective of your project? What outcomes do you expect after completing your first year of your PhD studies?

The objective of my project is to develop a long-term sensor for measuring neurotransmitter levels, which will be integrated with brain organoid(s) to model Parkinson’s disease. While commonly used electrodes quickly lose their sensing ability due to biofouling or electrochemical fouling, my aim is to produce functionalized carbon nanomaterial-based electrodes that preserve their ability to accurately measure neurotransmitter concentrations over months. After spending one year researching the subject, I expect having finished mechanical biocompatibility study of our electrode materials while also overcoming the biofouling problem.

Brain modelling elicits the idea of using computer simulations to model information flow in the brain, although in CONNECT project’s context it means using brain organoids to model functioning of the brain as a biological entity. Placing brain organoids onto microfluidic chip enables direct and precise control for administrating nutrients and drugs, which has already been introduced by other groups. Integrating this brain-on-a-chip with neurotransmitter sensor however would enable us continuous monitoring of the organoid’s condition in real-time.

I am currently not planning to attend conferences as my focus lies on completing experimental work required to publish in peer-reviewed journals. I believe this allows a more effective way of sharing research progress at this moment, rather than attending online conferences. Currently, my academic training also has a focus on educational activities, in which I lecture about the interaction between living cells and materials. I’m looking forward to attending conferences in the future when the COVID-19 situation eases out and face-to-face meetings in my field of research are organized again.

In your own words, how do you think does CONNECT make a difference to society?

It is difficult to say about the project’s rightful influence at this early moment, when the consortium is yet so young and we are still exploring various aspects of the highly interdisciplinary project. The already-obtained results from our individual consortium partners however suggest that CONNECT will have a significant impact on the development of organoid models, organ-on-chip technologies, and also electrodes for long-term interfacing with the nervous system. The first two allow for paradigm shift for drug development and thus decrease the percentage of failed human trials while simultaneously reduce the need for animal models. The technologies and materials developed for our integrated electrodes could also find use in applications that require an interface between computer and brain.

Is there something else you would like to share?

Interdisciplinary projects are highly enlightening because they require a certain kind of openness but also an interest in crossing the boundaries of one’s scientific bubble. I only wish such attitude would be more prevalent in the world we live in, especially from the non-scientific perspective. After all, we are currently having various societal and global problems that could greatly benefit from it.

Interview with Antigoni Gogolou, PhD student in CONNECT Nervous-system-on-Chip

Miss Antigoni Gogolou is a PhD student in the Tsakiridis Lab at University of Sheffield working on cell fate specification using human pluripotent stem cell- derived neural crest cells with a focus on the development of the enteric nervous system.Her scientific academic career started with her 5-year Degree (integrated Masters) in Applied Biology from the University of Ioannina in Greece, where she graduated in the top 3% of the entire course. During the final year of her studies, she carried out her diploma thesis project in the laboratory of Androniki Kretsovali at the Institute of Molecular Biology and Biotechnology in Greece, where she worked on the study of early embryonic development using mouse embryonic stem cells. In 2018, Antigoni decided to expand her knowledge on human embryonic stem cell biology through a 5 months internship in the Tsakiridis laboratory at the University of Sheffield, after receiving an Erasmus+ grant. Her interest in developmental biology, led her to pursue a PhD in the same group and her current work focuses on enteric nervous system development through the use of human pluripotent stem cells (hPSCs).

What is the objective of your project? What outcomes do you expect after completing your second year of your PhD studies?

The aim of the project is to construct a physiologically relevant enteric nervous system on a dish, that contains the appropriate enteric neural and glial components in content and at the correct numbers. By integrating our knowledge in hPSC differentiation with developmental biology, we aim to deliver an easy to follow and reproducible protocol to generate enteric neurons and glia, that can be used to study enteric nervous system (ENS) development and disease. Our ultimate goal, in the framework of CONNECT, is to combine hPSC-derived peripheral nervous system elements such as peripheral nerves and enteric neurons with brain organoids, developed by our partners, on a single smart chip to study the connectivity between different parts of the nervous system, a factor which has been found to play a profound role in neurodegenerative disorders such as Parkinson’s disease. We are currently in the process of characterising the hPSC-derived ENS components and towards the end of the second year of my PhD, I set my sights on having an optimised protocol developed.

What comes to your mind when you hear chip technology and why do you think it makes a difference to apply chip technology to enteric neuron biology?

As everyone else who is not an expert in microelectronics, the first thing that crosses my mind when I hear chip technology is the conventional computer chip with all the integrated microcircuits. However, in our case we are talking about sophisticated and elegantly designed micro-fluidic cell culture devices, that support cell growth and allow experiments to be done on living cells by providing stable conditions that mimic the natural environment. Luckily, chip technology and more specifically organ-on-chip technology, has attracted a great amount of interest nowadays, so that more and more people have become familiar with it. Combining chip technology with our living cells will not only add complexity and the required three-dimensional aspect to our system but will facilitate controlling and monitoring the cellular microenvironment, the major source of variability in cell culture. The application of chip technology to enteric neuron biology is likely to provide a more accurate model of human ENS development, surpassing the need of traditional animal models and the ethical issues associated with their use.

Are you planning to attend a conference presenting your research results soon? If yes, please tell us a bit more about the type of conference and who the main attendees are? Are there also participants from industry, representatives of patient groups or medical specialists?

I was planning to attend and give a poster presentation in Gastronauts, a symposium taking place in Nantes in May 2020, however, due to COVID-19 pandemic, unfortunately the organisers decided to postpone it for May 2021. Gastronauts Nantes is a symposium on gut-brain axis matters with a full agenda of exciting and multidisciplinary research from gut neurobiology to synthetic biology and bioengineering, primarily led by scientists from an academic environment. The attendee list includes students, post-docs, PIs, people from industry and in general, anyone who is interested in gut biology of a broader scope and is keen to share their enthusiasm with peers in the field.

In your own words, how do you think does CONNECT make a difference to society?

CONNECT’s holistic approach by integrating fully connected cultured organ systems such as central, peripheral and enteric nervous system provides a better insight into human physiology and offers a highly interconnected complex system that precisely emulates in vivo function from cell to organ level. Developing an in vitro full organ system like the CONNECT platform, will be highly beneficial to society for ethical and practical reasons. The CONNECT project will not only provide a tractable and reproducible system to model disease development and progression, but will also advance drug discovery by enabling high-throughput screening. Moreover, I believe that CONNECT’s novel approach will open up new venues for personalised medicine and the development of treatments tailored to patient’s health needs by using drug candidates on patient derived cells.

Interview with Yagmur Demircan Yalçin, Postdoc in CONNECT Nervous-system-on-Chip

Dr. Demircan Yalcin is a postdoctoral fellow of Neuro-Nanoscale Engineering at the Microsystems section in the Department of Mechanical Engineering, Eindhoven University of Technology, The Netherlands. She is working on the design and implementation of novel micro physiological sensors, which combine electrical and mechanical modalities. By using her hands-on experience with 3D electrodes in analyzing the biological cells in microfluidics applications, she focuses with her current research on “robustly re-using 3D MEAs in nervous-systems-on-a-chip: novel platforms and best practices”

Dr. Demircan Yalcin received her, B.Sc., MSc, PhD in Electrical and Electronics Engineering from the Middle East Technical University, Ankara, Turkey in 2010, 2013 and 2018, respectively and she also has a Minor Degree in Biology from the same university. She received her PhD. on the topic: A Lab-on-a-chip System Integrating Dielectrophoretic Detection and Impedance Counting Units for Chemotherapy Guidance in Leukemia. Before joining Neuro-Nanoscale Engineering at TU/e in January 2020, she worked at Mikro Biyosistemler, Ankara, Turkey as an R&D Engineer from 2015 to 2019.

What is the objective of your postdoctoral studies in CONNECT?

I am enthusiastic in combining engineering with biology. I already had the opportunity to be included in the development of a pioneering Lab-on-a-Chip diagnostic platform to provide better healthcare for people since I started my research life as a Master student 10 years ago. I observed that the creation of engineering tools to discover and understand biological mechanisms takes place at the interface of disciplines. In the CONNECT project, I believe this knowledge can make a difference because my scientific objective for CONNECT is the design and fabrication of an electrophysiological readout method being integrated with the microfluidic 3D cell culture environment of the novel Nervous-system-on-Chip. The CONNECT platform incorporates knowledge from lots of scientists from different areas. My personal objective for CONNECT is to act as a bridge between engineers and biologists by using my interdisciplinary background, including theoretical and practical experiences.

What do you think (from the viewpoint of your respective scientific discipline) is the biggest challenge in combining chip technology with neurobiology?

Real-time and label-free monitoring is the main challenge in my opinion because finding of a cost-effective material which has suitable properties to provide a proper environment for living cells and to make high resolution monitoring possible is difficult.

If you were meeting an investor in an elevator, what would be your 15 seconds Message Map to pitch to this CONNECT stakeholder the importance of putting the next level in vitro model systems for the nervous system firmly on the map?

Research in CONNECT links the nervous system of the brain and the gut in one single chip to understand mechanisms behind neurodegenerative diseases like Parkinson. CONNECT is an organ on a chip platform based on novel findings in stem cell technology and advances in microfluidics. It aims to increase the success of drug discovery studies and reduce the need of animal tests.

Is there something else you want to add?

Especially for the development of novel healthcare technology, working together across different scientific disciplines and types of technology is key. The different approaches and terminologies used in these diverse fields of research make effective collaboration a real challenge. However, it becomes really interesting and enjoyable after starting to understand the language of others. CONNECT literally connects us to be open minded in evaluating our findings not only from our own but also from the perspective of other disciplines including the neurosciences, which urgently seeks to apply nervous-systems-on-chip technology to advance their frontiers.

Interview with Gemma Gomez Giro, Postdoc in CONNECT Nervous-system-on-Chip

Dr. Gemma Gomez-Giro is a postdoctoral researcher at the Luxembourg Centre for Systems Biomedicine of the University of Luxembourg in the Developmental & Cellular Biology group (Schwamborn Lab).

Dr. Gemma Gomez-Giro received her B.Sc. in Biomedicine from the Autònoma University in Barcelona, Spain, in 2013 and continued with her master studies at the Pompeu Fabra University in the same region. After completing her MSc degree she moved to Munich, Germany, with an Erasmus+ grant pursuing her wish to investigate neurodegenerative diseases. Following her interest in neurodegenerative disorders, in 2015 she started her PhD in Biology at the Westfälische Wilhelms-Universität in Münster, Germany. She received her degree in 2019 on the topic: Modelling Juvenile Neuronal Ceroid Lipofuscinosis by genome editing in human induced pluripotent stem cells and cerebral organoids.

What is the objective of your postdoctoral studies in CONNECT?

The objective of my postdoctoral studies in CONNECT is to help integrate the knowledge we have gathered in the Schwamborn lab concerning neurodevelopment and 3D organoid culture technology in order to achieve a functional connection between different elements of the nervous system. It is my goal to participate in the production and up-scaling of the central nervous system compartment, which should contain all the necessary elements to establish a network with the other elements on the chip. It is also my objective to work on the application of the system to study Parkinson’s disease.

What do you think (from the viewpoint of your respective scientific discipline) is the biggest challenge in combining chip technology with neurobiology?

In my opinion, the random configuration of organoid 3D cultures makes it difficult to precisely control their growth and directionality. However, on-chip platforms are already developing better tools to achieve more guidance and monitoring of the microenvironment, overcoming at the same time concerns regarding reproducibility. I believe the biggest challenge when bringing both aspects together is the need to incorporate multi-omics readouts and advanced imaging techniques to be able to capture interactions and physiological responses at the systemic level and gather high-throughput data from the microfluidic system. Necessarily, this has to be paired with advances in computational models that can support such data analysis in order to achieve faster and reliable outcomes.

If you were meeting an investor in an elevator, what would be your 15 seconds Message Map to pitch to this CONNECT stakeholder the importance of putting the next level in vitro model systems for the nervous system firmly on the map?

In the CONNECT project we believe that it is not possible to explain complex neurodegenerative diseases by looking at the different systems affected separately. Therefore, we aim to increase the complexity level of in vitro systems to establish a functional connection between different elements of the nervous system. In order to do that, the advances in on-chip technologies are crucial in helping us establish, examine and measure this interaction. CONNECT represents an important bridge from the bench to translational medicine, allowing us to gain insight in Parkinson’s disease pathology mechanisms. Moreover, CONNECT is an innovative, improved and attractive in vitro screening platform that could reduce time and cost of drug development and testing, replacing animal experimentation.