LI Blog: Yvonne, welcome to the Yokogawa Life Science Europe Team! Could you please introduce yourself briefly—your background, current role, and what brought you to Yokogawa?

Dr. Yvonne Dürnberger: Thank you. I’m very excited to join the Yokogawa Life Science Europe Team as Business Development Manager. I have a background in biology and biomedicine, and I was fortunate to work with the Cell Voyager 6000 during my PhD studies at the DZNE in Bonn. Before I joined Yokogawa, I worked for five years as a team leader in the product management and marketing department of a biotech company, where I was responsible for launching and managing the life cycle of a high-content imaging platform. At some point, I realized it was time to follow a different dream for a while. So, my partner and I quit our jobs and traveled the world for one and a half years. During that time, we were fortunate enough to experience new countries, new cultures, marvel at new wonders, and go on countless adventures. Now, I feel the circle is closing in on itself, bringing me back to where I originally started – Yokogawa’s high-content imaging world.

Supporting Co-Innovation and Strategic Partnerships

LI Blog: In your role as a Business Development Manager, what are your core responsibilities, and how do they contribute to Yokogawa’s mission?

Dr. Yvonne Dürnberger: In my role as Business Development Manager within the Life Science Europe Team, my responsibilities are quite diverse. I work closely with our high-content imaging distribution partners and help further develop both existing and new strategic partnerships in support of Yokogawa’s mission of co-innovating tomorrow. In addition, I also support the team in various areas such as application, promotion, and marketing.

How Confocal Imaging Becomes More Accessible

LI Blog: What makes the CQ3000 stand out, and why do you believe it offers real value to our customers?

Dr. Yvonne Dürnberger: I still remember my first impression when I was introduced to the CQ3000: this device is going to be a game changer. I believe this system is the sweet spot between highly advanced high-content imaging flagships and affordable, compact confocal imagers. It comes in a bench-top design, is equipped with extremely fast high-resolution confocal and live-cell imaging capabilities, is easy to use, and is offered at a fair price. I am excited to see this instrument conquer the market.

LI Blog: What aspect of your work do you enjoy the most, and what keeps you motivated every day?

Dr. Yvonne Dürnberger: I love to work in exciting fields that are constantly met by new challenges, learn to evolve, and are fueled by innovation. And if you share your workdays with a group of very capable, highly supportive, and extremely kind individuals, this is where I find my motivation.

The Beauty and Science of Cell Imaging

LI Blog: What sparked your interest in imaging, and how has that passion evolved?

Dr. Yvonne Dürnberger: Microscopy and different variants of high-content imaging have accompanied me throughout my professional life, from my early studies to my new role here now at Yokogawa. From the very start, I have always been drawn to beautiful cell or tissue images and, early on, realized that these images themselves are not the end but only the beginning of finding the answers to your scientific questions. Having faced the challenges of imaging and data analysis myself as a young scientist and now seeing how these technologies have evolved in such a short amount of time is incredible. I am very much looking forward to witnessing how this progress continues in the future.

Work Life at Yokogawa

LI Blog: What have you enjoyed most so far about working at Yokogawa?

Dr. Yvonne Dürnberger: From the very first day, everyone at Yokogawa, and specifically the Life Science Team members, has been incredibly welcoming and supportive. This feeling of being welcome in addition to continuously learning more and more about these great technologies, is something I have enjoyed very much.

LI Blog: Is there something you’d like to share with our blog readers?

Dr. Yvonne Dürnberger: If you ever want to know the best spots for seeing a whale jumping out of the water, a turtle laying eggs in the middle of the night, or witnessing a volcano spitting lava while standing on the summit of a second volcano, I am your contact 😉

The post High-Content Imaging Meets High Adventure – An Interview with Yvonne Dürnberger appeared first on Life Innovation Blog.

Eric Sobierajski: Thanks for the warm welcome. My name is Eric Sobierajski, and I have recently joined the Life Innovation Business Team as a product manager for the SCA and CSU products. Before I started at Yokogawa, I was a research associate at the Ruhr University in Bochum, where I also graduated in Neurobiology. During my doctoral thesis, I frequently used confocal microscopy to image and analyze different cellular components and their development in the brain. The passion to work with this fascinating kind of devices and the possibilities that they are giving us in the perspective of scientific research led me to this position at Yokogawa.

What It Means To Be a Product Manager

LI Blog: In your role as a Product Manager, what are your core responsibilities, and how do they contribute to Yokogawa’s mission?

Eric Sobierajski: The tasks of a Product Manager consist of a repertoire of responsibilities. Leading the products to their desired purposes, this requires close cooperation with customers, sales, marketing and production. Market analysis for potential competitors as well as requirement analysis are also on the list of tasks. In doing so, we aim to achieve Yokogawa’s mission, providing the most reliable incubation and live-cell imaging systems to the world, framed with excellent service.

Innovating with Impact

LI Blog: What makes the SU10 stand out, and why do you believe it offers real value to our customers?

Eric Sobierajski: The Single Cellome^TM Unit 10 offers a fully automated and targeted injection of a variety of compounds like biomolecules, drugs, and gene editing tools (CRISPR/Cas9) and offers therefore a great variability to influence single cells or even cell organelles. What makes the SU10 unique is its high injection success rate and cell survival rate because it is minimal invasive. Moreover, the SU10 is easy to use because deep knowledge of microscopy is not required. Knowing the struggle from my doctoral period, these aspects are really of high value.

What Fuels Eric’s Daily Motivation

LI Blog: What aspect of your work do you enjoy the most, and what keeps you motivated every day?

Eric Sobierajski: I like the fact that the work as a product manager gives you insights into the different departments of the company and that I can learn new things “outside the box” every day. Since we are a small team, I have also the opportunity to choose parts of my tasks I want to focus on in particular. And finally, not losing the connection to science is very important for me.

Why Neurobiology?

LI Blog: What sparked your interest in neurobiology, and how has that passion evolved over time?

Eric Sobierajski: Since I can actively remember (and according to my parents also the time before), I always loved watching animals and plants in nature and going to the zoo. With the years more and more questions around the “why’s” and “how’s” rose my curiosity, which hardened during school and afterwards. Neurobiology – “the brain thinking about itself” – was my favorite discipline after a short time. And after years of study in which I learned a world about the nervous system, there’s still worlds that we just begin to understand.

Life and Culture at Yokogawa

LI Blog: What have you enjoyed most so far about working at Yokogawa?

Eric Sobierajski: At Yokogawa, the spirit of the people working here is wonderful and I really like the relaxed and friendly atmosphere and working culture. So, this made my start easy, and I look forward to the challenges that are waiting for the Life Science Team.

LI Blog: Is there something you’d like to share with our blog readers?

Eric Sobierajski: I am always happy to talk with other people who have the same fascination about the today’s possibilities in microscopy and confocal imaging!

The post A Passion for Microscopy and Innovation: Interview with Eric Sobierajski appeared first on Life Innovation Blog.

What is Blue Carbon?

Blue carbon refers to the carbon stored in coastal blue carbon ecosystems such as salt marshes, seagrasses, and mangroves. These ecosystems act as vital carbon sinks, sequestering large amounts of carbon dioxide from the atmosphere.

Unfortunately, these ecosystems have been disappearing at an alarming rate. Over the past several decades, an estimated one-third of global coastal blue carbon ecosystems have been lost. This destruction has dire consequences, releasing between 0.15 and 1.02 billion tons of carbon into the atmosphere annually, thereby contributing to climate change.

The Role of Blue Carbon Ecosystems

Coastal blue carbon ecosystems are invaluable not only for sequestering carbon but also for their role in maintaining biodiversity and supporting human wellbeing. However, their degradation turns them from carbon sinks into carbon sources, intensifying global warming and disrupting marine life.

The Impact of Global Warming on Marine Ecosystems

One example of how global warming affects ecosystems is the rapid growth of green seaweed Ulva, especially in coastal regions. For instance, Ulva prolifera (U. prolifera) has become the dominant species in the Yellow Sea in China, creating trans-regional disasters through macroalgal blooms known as green tides.

Key Factors Influencing Ulva Growth

The proliferation of U. prolifera typically begins in the southern Yellow Sea during mid-April to early May, with seawater temperature identified as a crucial factor. At the end of green tides (mid-July to early August), massive fronds of U. prolifera sink to the seabed or remain suspended at varying depths, where they gradually decompose.

Image Credits: 123RF / Sosiukin

Environmental Consequences of Green Tides

The decomposition of U. prolifera has profound effects on coastal ecosystems:

• Oxygen depletion: Microorganisms breaking down the fronds consume large amounts of oxygen, leading to low-oxygen or hypoxic conditions.
• Marine life deaths: Hypoxia can result in the mass death of marine organisms, particularly in aquaculture zones.
• Carbon source creation: Coastal waters become carbon sources as large amounts of CO2 dissolve into seawater during decomposition. However, this process also contributes to the long-term accumulation of organic carbon, aiding oceanic carbon sequestration.

Exploring the Positive Potential of U. prolifera

Despite its detrimental environmental impact, U. prolifera’s rapid growth offers opportunities for positive applications, including its use in biofuels and bioremediation.

Advancing Research on U. prolifera with Genome Editing

The research group led by Kensuke Ichihara (Phycological Research, 2022) has been investigating the genomic functions of U. prolifera. Genome editing techniques, particularly the CRISPR-Cas9 system, have been instrumental in uncovering the genetic mechanisms behind U. prolifera’s proliferation.

Genome editing is a powerful tool to understand gene function by causing a specific mutation on the target gene and was used by Kensuke Ichihara et al. The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system is a simple genome-editing system based on bacterial immunity. The delivery of gene-editing materials like CRISPR-Cas9 can be performed by our Single Cellome™ Unit SU10. The genome-editing methods can broaden the scope of functional genomic studies of Ulva.

Please visit our website for more information: https://www.yokogawa.com/eu/solutions/products-and-services/life-science/single-cellome/su10/

The post Blue Carbon – A Nature-based Solution for the Climate, Biodiversity and Human Wellbeing appeared first on Life Innovation Blog.

Daisuke Nojima: Thank you all for the warm welcome. I am deeply honored to take on the role of Managing Director at Yokogawa Innovation Switzerland (YIS). I recognize there is much to learn in this new position, and I sincerely appreciate the ongoing support of Yokogawa colleagues in Europe as we work together to advance YIS.

Our goal is to significantly strengthen YIS’s presence in Switzerland and across Europe, with a particular focus on biotechnology and life sciences, which are key growth areas for Yokogawa. In 2024, we aim to assess the business potential of our current biomanufacturing project. YIS will continue to build collaborative relationships with internal teams and external partners, such as Yokogawa’s Bio Center of Excellence.

I look forward to working with all of you as we drive innovation and expand our capabilities in these exciting fields.

LI Blog: What services do you offer in your shared laboratory at Switzerland Innovation Park Basel?

Daisuke Nojima: Our laboratory at the Switzerland Innovation Park Basel is equipped with a range of Yokogawa’s advanced life science products, which we use for our R&D initiatives. The facility also serves as a showroom, where we showcase Yokogawa’s cutting-edge solutions in life sciences and conduct live demonstrations for our customers. Notably, it is the only Yokogawa laboratory in Europe capable of performing bio experiments at the BSL2 (Biosafety Level 2) standard, enabling us to conduct sophisticated biotechnological research and development on-site.

Wanted: Partners with Expertise in Single-Cell Manipulation

LI Blog: Which companies or research institutes would you like to cooperate with?

Daisuke Nojima: As part of Yokogawa’s commitment to advancing the bioeconomy, YIS plays a key role in fostering innovation in this space. Through open innovation and an awareness of evolving social and environmental needs, YIS remains open to collaboration with companies and research institutes across various sectors.

For our current project, we are actively seeking partners with expertise in single-cell manipulation technology, particularly for microalgae, as well as downstream biomanufacturing processes such as advanced cultivation technologies. We are eager to explore opportunities that can accelerate innovation in these areas and enhance our capabilities in biomanufacturing.

LI Blog: Could you provide an update on the development of technologies for smart cell creation with Yokogawa Technology Solutions India (YTI)? What is the current status?

Daisuke Nojima: Our project, which began in 2021, has made significant progress. At YIS, we have successfully developed single-cell manipulation technology for microalgae, while YTI has focused on creating bioinformatics and automation software for metabolic and other genetic engineering target predictions to support biomanufacturing processes. We are now in the process of integrating these two technologies into a unified system aimed at establishing smart cell development. Additionally, we are preparing to evaluate the business feasibility of this initiative in collaboration with external partners, moving closer to practical applications.

LI Blog: What has been your most enormous success so far?

Daisuke Nojima: Our most considerable success has been raising awareness of YIS across Europe. When YIS was first established in Basel in 2020, few people, even within the Yokogawa group, were familiar with us. Thanks to the strong support from the Embassy of Switzerland in Japan and Switzerland Innovation, we have enhanced our visibility greatly. They provided valuable opportunities to connect with influential individuals and participate in high-profile events, helping us establish a foothold in Switzerland’s innovation ecosystem.

Additionally, JETRO (Japan External Trade Organization) has been instrumental in expanding our network throughout Europe. Through events they organized, we’ve had the opportunity to meet and engage with Japanese companies in Europe, many of which share a similar mission. These conversations have been both encouraging and inspiring. JETRO has also facilitated introductions with top-tier companies and universities, allowing us to share ideas and explore collaborative opportunities.

Through the combined efforts of the Swiss Embassy, Switzerland Innovation, and JETRO, YIS has built a robust network of contacts that will be vital for future collaborations. We are deeply grateful for their continued support.

Next Trends in Biotechnology

LI Blog: What do you think will be the next trends in biotechnology?

Daisuke Nojima: I believe that the drive to reduce CO2 emissions will be a major force propelling advancements in biotechnology. The European Commission, for example, has adopted ambitious proposals to reduce net greenhouse gas emissions by at least 55% by 2030, compared to 1990 levels, and many other countries are following suit with similar initiatives.

One area with significant potential is the development of materials derived from biomass and bioprocesses, although the high costs currently present a challenge for widespread adoption. However, as regulations tighten and industries are required to shift towards more sustainable alternatives, even at a higher cost, biomanufacturing will emerge as a crucial solution. I am closely monitoring these regulatory trends, as they will undoubtedly shape the future direction of multiple industries.

Cultural Differences

LI Blog: After living in Switzerland for almost 4 years, what is the biggest difference compared to Japan?

Daisuke Nojima: The cultural differences between Europe and Japan are significant, particularly in areas like work style and communication. It can be challenging to adapt if one strictly adheres to traditional Japanese approaches. However, this challenge is not unique to Europe; it highlights the importance of continuously evolving personal and professional habits to succeed and maintain performance across different cultural contexts.

Fortunately, through open communication with colleagues in Europe and interaction with other expatriates, I’ve been able to adjust my mindset and adapt to the unique dynamics of working in Switzerland.

LI Blog: Where can our readers meet you? What will be the next event you’re attending?

Daisuke Nojima: I will be attending upcoming pitching events organized by JETRO and several conferences focused on biomanufacturing. I welcome anyone interested in connecting to contact me via LinkedIn or email. I’m always excited to meet new people and explore shared interests. Networking and exchanging ideas are crucial to driving innovation and achieving success, and I look forward to engaging with you all.

For more information about YIS, please visit their website.

The post Biomanufacturing will emerge as a Crucial Solution – Interview with Daisuke Nojima appeared first on Life Innovation Blog.

Enhanced Features for Single-Cell Analysis

This July, Yokogawa Deutschland GmbH is excited to introduce the updated SU10, which comes with several enhanced features designed to improve user experience and performance:

•  Enhanced Motor Actuator Robustness: The motor actuator has been fortified for greater durability, ensuring more reliable and consistent results.
• Improved Position Repeatability: The SU10 offers more precise nanopoint delivery, enabling accurate positioning inside the cell.
• Flexible Manipulator Movement: In contrast to the previous model, the manipulator can be moved. The condenser lens can still be used when SU10A is not in use.
• Compact Design: The controller boxes are reduced from two boxes to one box.

Nanopipette Technology

The Single Cellome SU10 is a cutting-edge solution for single-cell analysis. Using nanopipette technology facilitates the delivery of different substances into the cytoplasm or nucleus of single cells in a semi-automated manner. It can also perform sampling at the single-cell level. Initially developed in the lab of Prof. Nader Pourmand at the University of California, Santa Cruz, the system has been further advanced by Yokogawa. For more details, please view our on-demand webinar Nanopipette Technology – A new Tool for Single-Cell Analysis.

The post New SU10 Model Release appeared first on Life Innovation Blog.

What you will learn in this on-demand webinar:

• The challenges associated with generating usable genome-modified clonal cell populations.
• How nanopipette injection can cut clonal cell population generation times to as little as ~3 weeks.
• Ways to utilize nanopipette injection in a small biotech setting.

Genome modification has become essential to most Biotechs’ research and development pipelines to produce new cell lines that convey a modification, such as a fluorescently labeled target protein, knock out of a target, or mutation of a target. 

Two general approaches are commonly used: electroporation of nuclease and nucleic acids (i.e., Cas9 RNPs) or transfection of vectors expressing nucleases, targeting nucleic acids and donor genetic material. 

One of the significant limitations of these approaches is that they take a long time to generate a usable clonal cell population due to the need for selection and population expansion, usually with some type of cell sorting. This process can often take several months.

In addition, electroporation or transfection does not work for some cell types, especially when inserting large DNA fragments such as genes encoding full-length fluorescent proteins, where correct insertion rates can drop below 0.01%. 

Nanopipette injection offers an alternative method of delivering genome-editing material, such as Cas9 RNPs with donor DNA. A major advantage of Yokogawa’s SU10 nanopipette injection system is the ability to efficiently genome edit cells at a single-cell level, allowing the user to start with a clonal population. The nanopipette injection system can help generate cell lines in as little as ~3 weeks. In this webinar, you will hear my experience of the SU10 system, including my impressions and my opinion on how small biotechs can best utilize it.

Single Cellome Unit SU10 Update

The SU10 recently received an update. For more details, please visit the SU10 product website.

The post On-Demand Webinar: Improving Genome Editing Pipeline Efficiencies with Nanopipette Delivery appeared first on Life Innovation Blog.

Stephan Müller: “I’m Stephan, recently appointed as a Field Service Engineer at Yokogawa’s Life Science division. Before joining Yokogawa, I worked in academia. As a former Drosophilist (Drosophila melanogaster, or commonly known as “the fruit fly”), I have explored different facets of this fascinating animal from an imaging standpoint. Drosophila researchers often make great microscopists, too!

In my (non-)professional life I like to understand how things work and to fix them. Or at least I’ll try. I just like to see that stuff can be re-purposed.  If there is time left, I like traveling, preferentially on a motorbike.”

The Core Role of a Field Service Engineer

LI Blog: What are your main tasks as a field service engineer?

Stephan Müller: “The job evolves around all aspects of a product life cycle when, or shortly before, being in the hands of the end-user.  Fixing, explaining, and figuring out what’s wrong, Or, in almost all cases, right.”

Advantages of using the CellVoyager CV8000

LI Blog: Why should customers decide to buy the CellVoyager CV8000?

Stephan Müller: “As with motorbikes, Japanese microscopes stand for quality. During my live-imaging focused PhD, I would have loved to have access to a product such as the CV8000.  Scientists should have the time to keep their minds on actual science, rather than laborious, yet repetitive, tasks. The CV8000 automatizes your entire imaging workflow while keeping your samples alive throughout the duration of your experiment. Such an automatically maintained experimental environment, in combination with gentle light exposure particularly suited for long-term experiments, would have saved me and those poor PhD students who are not yet done with their PhDs, months, if not years, of a valuable lifetime, spent dealing with equipment that isn’t properly functioning.”

Passion for Live-Cell Imaging

LI Blog: What do you like most about your job?

Stephan Müller: “I’m now getting paid for that part of my previous job that I liked the most but has been unpaid. It is fascinating to work and understand the very devices that are used to describe as accurately as possible what nature had millions of years to perfect. It’s just great to see that we are nowadays able to observe life well beyond the extend of what I have dreamed possible ten years ago. And it will only get better.”

LI Blog: Why did you decide to focus on live imaging?

Stephan Müller: “For decades, researchers have been able to resolve fine details of whatever specimen using techniques such as electron microscopy. However, fixed samples are, as the names suggest, fixed or inherently dead, respectively. Being alive is the exact opposite of a fixed sample. So understanding cellular dynamics is a task that can only be performed on living samples. The 20^th century was about structure, the 21^st century will be about dynamics. Biology will become the most data-intensive scientific research area, and it’s just very cool to be part of it.”

Academia vs. Corporate

LI Blog: You worked at a university research institute before. If you compare, what is the biggest difference between an academic position and a corporate job?

Stephan Müller: “As of now, it doesn’t feel overly different. The actual work seems very similar, it’s just that the working conditions are so much better outside academia. Additionally, the corporate world relies on earning its own money. Unlike academia, where funding is always measured by the amount that comes in, rather than the use of it, in the corporate world, spending is differently justified. It seems to me as if this makes processes way more efficient rather than relying on more working hours of (mostly underpaid) scientific personnel.”

The Yokogawa Spirit

LI Blog: What do you like most about working at Yokogawa?

Stephan Müller: “From what I’ve seen so far, there seems to be a clear distinction between work and life, while keeping the professional life flexible enough to fit to nowadays adult lives.  Also, people seem to have very clear job descriptions, there is always someone who knows what to do or whom to ask.”

A personal Message to our Readers

LI Blog: Would you like to share something with our blog readers?

Stephan Müller: “Ask questions and happy imaging!”

The post Imaging and Engaging with Microscopes is incredibly rewarding! -Interview with Stephan Müller appeared first on Life Innovation Blog.

LI Blog: Could you tell us about your academic journey and how you developed a keen interest in microscopy and quantitative imaging?

Sven Fengler: Absolutely! I am a biologist by training. My academic journey began with my undergraduate studies at the University of Cologne. Conducting quantitative FRET experiments by acceptor photobleaching allowed me to explore protein-protein interactions and caspase activity, particularly in the context of the NF-κB-inflammation signaling cascade, which involved a myriad of proteins. This experience ignited my passion for using advanced imaging techniques to study intrinsic biological processes.

LI Blog: How did your Ph.D. research contribute to your expertise in microscopy?

Sven Fengler: My time at the Max Planck Institute in Dortmund, working on my Ph.D., was transformative. Particularly combining FRET-FLIM assays with automated microscopy, allowed me to explore protein phosphorylation events inside cells with spatial resolution in a high-content format. I used this technique to investigate the phosphorylation status of the EGF-receptor and its regulation by phosphatases during receptor signaling and recycling events. I developed reverse transfection cell arrays that allowed me to express 384 different proteins from cDNA libraries on a miniaturized format with a size of a microscope slide. Part of my work was to automate the cell array production for my PhD project.

LI Blog: How did working with physicists influence your work?

Sven Fengler: It was inspiring to work together with physicists in the team because it allowed me to gain a deep understanding of the technical aspects in laser optics and quantitative image analysis. In particular, working in a team of experts to develop an automated microscopy setup to image transfection cell arrays with FRET-FLIM optics was groundbreaking for me.

LI Blog: As an early PostDoc, you transitioned to DZNE Bonn. Could you tell us about your work there?

At the DZNE Bonn, I found the perfect match for my skills and interests. Therefore, I joined the team of Laboratory Automation Technologies (LAT) within the Core Research Facilities and Services (CRFS) in 2017. My work focuses on the assay development of advanced human iPSC-derived cell models for complex 3D systems and preclinical drug screenings. The dynamic research environment at DZNE Bonn has allowed me to grow professionally and personally.

Research Focus

LI Blog: What is your main research focus?

Sven Fengler: Recently, we have established a novel blood-brain barrier (BBB) system, which I have also presented at the CV8000 User Group Meeting in 2022. The constructed blood-brain barrier endothelium of the system derives from human iPS cells. In the past years, we developed a 3D microvessel system, like a small blood vessel in vitro. The aim was to establish a screenable system that mimics the complex architecture of the human BBB as close as possible to provide a technology to screen for BBB-penetrating substances early – in the phase of preclinical drug discovery.

The Importance of the Blood-Brain Barrier

LI Blog: What is the blood-brain barrier (BBB), and why is it important to study the BBB?

Sven Fengler: This specialized endothelial structure, present in all of us, plays a crucial role in maintaining healthy brain function and preserving the brain’s delicate internal environment. This selective permeable barrier allows essential nutrients and molecules to enter the brain while keeping toxic substances at bay. The key players in this defense are efflux transporters, which actively pump out harmful substances back to the blood cycle. By doing so, these transporters safeguard the brain’s delicate microenvironment, ensuring it remains free from detrimental agents that could disrupt neurological homeostasis. However, when you think about designing a drug against Parkinson or Alzheimer diseases for example, you need to overcome this barrier and “trick” these transporters, the endothelium per se. Overcoming this challenge is crucial for the success of potential treatments in general. The high failure rate of drug candidates in clinical trials can often be attributed to their inability to penetrate the BBB effectively.

Recent iPS cell technology has the potential to mimic the complex architecture of the human BBB much closer than it was possible a couple of years ago. Such systems could help us to test and finally better predict a BBB-penetration of a new drug.

The Drug Discovery Process for Neurodegenerative Diseases

LI Blog: Could you explain to us the conventional drug discovery process for neurodegenerative diseases?

Sven Fengler: During lead optimization, researchers employ a variety of experiments to predict the drug’s ability to cross the BBB. Often, these assays use artificial membranes or Transwells, which may not accurately replicate the physiological conditions of the human BBB. For example, Transwells experiments typically comprised of non-brain tissue cell lines, might not exactly mimic the relevant properties of the very complex BBB system present in the human brain.

LI Blog: So, there are limits in preclinical testings?

Sven Fengler: Exactly. When potential drug candidates fulfill a consortium of properties measured in various in vitro assays, researchers assess whether a drug can penetrate the mouse brain to generate relevant in vivo data. However, optimizing lead-compounds based on mouse BBB permeability might not yield results that accurately represent the human BBB’s behavior. Consequently, many drug candidates may fail later during clinical phases due to their inability to cross the human BBB effectively.

LI Blog: Why could iPS cells overcome these hurdles?

Sven Fengler: We are hopeful that iPSCs can offer a more relevant and human-specific approach to address BBB permeability concerns. iPSCs can be generated from human tissues and have properties that mimic human BBB physiology better than traditional approaches. By using iPSCs to create endothelial cells that closely resemble the physiological brain endothelium, scientists can establish a more accurate model of the BBB. iPSC-based endothelium offers a regenerative system that can be grown for extended periods while maintaining characteristics closer to primary human cells. This advantage is particularly relevant for testing substances over a long experimental time. By measuring the Trans-Endothelial Electrical Resistance (TEER) of these cells, researchers can assess the integrity of the endothelium, gauging whether it forms a robust and tight barrier, akin to the human BBB.

3D human iPSC-derived Blood-Brain Barrier System – Technical Details

LI Blog: What is the technical background of your 3D human iPSC-derived blood-brain barrier system on a chip?

Sven Fengler: In our endeavors to mimic the physiological state of the BBB, we have characterized tight-junction proteins expressed by endothelial cells. These proteins play a pivotal role in the “passive” barrier function of the BBB. By measuring the TEER of our 3D endothelial vessel, the barrier property can be assessed quantitatively. To that end, our generated endothelium in culture shows physiological relevant barrier properties by reaching TEER values that resembles those measured in rodent brain vessels. We could experimentally show that our 3D system mimics the complexity of BBB architecture, including polarized efflux transporter localization. Finally, we simulate a medium flow that closely resembles the blood flow in small blood vessels.

LI Blog: Do you use special plates for imaging?

Sven Fengler: By seeding our IPSC-derived endothelial cells into the microfluidics chip (OrganoPlate® 3-lane 40) from Mimetas, remarkably, the endothelium self-organized into a vessel within two days. The grown vessel exhibit wound-healing properties by keeping its mono-layer structure that requires perfusion for optimal functionality. Using Mimetas plates, the medium perfusion is applied by a simple gravity flow simulating the blood flow inside each vessel. Each plate can contain up to 40 microvessels, providing sample testing capacity. Additionally, the system is compatible with imaging, allowing us to observe and analyze the microvessels and their interactions with other brain cells.

LI Blog: How was the imaging further performed?

Sven Fengler: For the microvessel characterization as well as for the high-content analysis in our permeability assays, we’ve used the Yokogawa CellVoyager CV8000. This state-of-the-art system allows us to image the microvessels fully automated and obtain fast, stitched 3D stacks with different fluorescent channels. The speed and efficiency of this system have enabled us to generate a wealth of data. Due to the integrated incubator system, live-cell imaging conditions are excellent. In this context, we have used the CV8000 to visualize the BBB penetration of different fluorescent tracer molecules to test microvessel integrity and to screen for toxic substances.

LI Blog: Could your model potentially enhance the success rate in drug discovery by neglecting costs at the same time?

Sven Fengler: By simulating the physiological state in vitro, our model provides valuable insights into how novel compounds might interact with the human BBB before testing them in vivo or clinical trials. With our system, we could evaluate a wide range of drug candidates and identify those with a relevant human BBB penetration already during preclinical drug discovery phases and lead optimization. This might enable researchers to focus resources on the most promising candidates and increase the likelihood of successful clinical translation.

Ideal Users of the 3D human iPSC-derived Blood-Brain Barrier System on a Chip

LI Blog: Who would benefit the most from using your system?

Sven Fengler:

Our model’s scalability and imaging capabilities further enhance its utility, allowing us to generate a vast amount of data and create comprehensive 3D animations for analysis. Possible applications are not limited to drug discovery alone. Our model can be applied in many other research fields including immune cell migration, virus and capsid penetration or toxicology for example. A huge advantage would be less animal testing in general because our system could provide an alternative to animal models in future.

Ultimately, our vision is to leverage this technology to streamline drug development, reduce failures in late-stage clinical trials, and pave the way for more effective treatments for neurodegenerative disorders.

LI Blog: What did you investigate exactly with the system?

Sven Fengler: In particular, we have tested the BBB-penetration of a set of anti-inflammatory substances. It has been shown that inflammatory processes are involved in the progression of many neurodegenerative diseases. Therefore, we focused on the identification of BBB-penetrating anti-inflammatory compounds because this group of drugs could have a potential to cure or stop disease progression in various forms of dementia.

Outlook

LI Blog: You have already achieved a lot with the 3D blood-brain barrier system. What are your future research plans?

Sven Fengler:

I’d like to mimic a blood-brain barrier from patients with a neurodegenerative disease.  For this, reprogrammed iPSCs derived from patient blood cells with a genetic disease background could be differentiated and matured to endothelial microvessels. Such an in vitro system could be used to simulate the effect of a compromised BBB due to the given type of dementia. For example, drugs could penetrate the BBB much stronger in case the BBB is leaky due to a specific disease and free drug concentrations could strongly differ from healthy to disease individuals.

Recently, we have set up our automated iPSC production robotics platform to produce iPSC-derived cells under reproducible conditions. We established protocols to freeze fully differentiated ready-to-use brain endothelial cells for our current partners in-house and for upcoming external collaborations.

LI Blog: Your research could be a lifesaver. Thank you so much for sharing your journey with us. Your work at the intersection of biology and microscopy is truly inspiring! We wish you continued success in your research at DZNE Bonn.

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What are Lipids?

Lipids are organic compounds consistent with nonpolar molecules, which are soluble only in nonpolar solvents (e.g., chloroform) and insoluble in water because water is a polar molecule. In the human body, these molecules can be synthesized in the liver.  

Lipids are an essential part of a cell`s biomolecular pool and play important roles in a myriad of complex biological processes like:

• Supporting cells and aiding in essential functions
• Protecting nerve cells
• Helping the body absorb certain vitamins
• Helping produce hormones, including estrogen, testosterone, and cortisol
• Energy storage (in the form of fat)
• The structural component of the cells
• Signaling functions

Changes in lipid composition are significant in fundamental biological processes such as growth, development, differentiation, and aging and are also important in disease states such as cancer. Being able to detect and quantify lipid composition in single cells would provide enhanced data on such cell states and reveal new information on the cellular role(s) of lipid heterogeneity.

For example, cancer cells show a wide variety of lipid profiles at the single-cell level. Cancer cells are highly heterogeneous and very diverse in their phenotype which makes them hard to extinguish, but the analysis of single cells and the characterization of the heterogeneity could provide new treatment strategies.

Lipidomics: Analysis of Lipid Profiles

Lipidomics is a newly emerged discipline that studies cellular lipids on a large scale based on analytical chemistry principles and technological tools, particularly mass spectrometry (MS). Recently, techniques have greatly advanced, and novel applications of lipidomics in the biomedical sciences have emerged.

MS-based techniques for the analysis of lipids are different in the absence or presence of liquid chromatography (LC) prior to mass spectrometric analysis. They can also be different in their analytical coverage (e.g., “targeted” vs “global” analysis). 

LC is a separation technique for lipids in which the mobile phase is a liquid that can be carried out either in a column or a plane. 

Challenges for Analysis of Lipid Profiles

One of the challenges is the low abundance of analytes present in one cell. There are new methods emerging on the marked dealing with such challenges. Another challenge is the disturbance of the natural state of cells by current instruments like microfluidic systems or machines that are droplet-based. The cells lose their spatial context by being detached from the surface they are cultivated on. Also, minor changes to the environment, which stress the cells, can change their lipid profile.

The sensitivity also plays a pivotal role in single-cell lipidomic studies and is another challenge that needs to be addressed.

Single-Cell Lipidomics Performance after SS2000 Single-Cell Sampling

For the first time, researchers at Surrey University could, with the help of SS2000, combine live cell imaging of pancreatic cancer cells with single-cell lipidomics. The results show an increase in the lipid droplet size (figure C) which was detected based on fluorescence microscopy and found to correlate with the higher abundance of neutral DG (Diglyceride) and TG (Triglyceride) lipids after induced oxidative stress (figure D).

This novel methodology has demonstrated unique capabilities. These were otherwise inaccessible to mass spectrometry imaging because living, single cells were sampled, and fluorescent live cell imaging was used on the same cell culture from which cells were sampled.

To demonstrate the capability of the developed lipidomics method for biological applications, the researchers sampled living single cells from two different cell cultures (PANC-1 and THP-1) and analyzed their lipid profiles. The results showed that the two groups separated significantly (figure A). This means that different cell types can be distinguished based on their lipidome.

The success of this methodology has been demonstrated in its ability to distinguish different cell types based on their lipidome (figure A) and make single-cell lipidomic observations consistent with previous observations in the literature based on cell extract analysis (figure B-D).

Why study Cancer by Single-Cell Lipidomics?

Lipids play many key roles in all basic processes essential for tumor development:

1. Lipids affect cell growth and metabolism, which are essential for rapidly proliferating cancer cells as the major building blocks for lipid biosynthesis and remodeling:

• Lipids represent the major structural components of cellular membranes (cholesterol, phospholipids, and sphingolipids)
• Lipids serve as the energy storage depot (triglycerides)
• Lipids are involved in energy metabolism and ATP production (acyl CoA and acylcarnitine)

2. Lipids are important for cell signaling:

• Lipids function as second messengers and as hormones in cancer cells to promote cell proliferation, survival, and migration (bioactive lipids, such as lysophospholipids)

Lipids are involved in significant cell signaling processes in chemotherapy and radiotherapy for human cancers.

We wish Dr. Melanie Bailey, Dr. Johanna von Gerichten, and Kyle D. G. Saunders a lot of success with their further research.

Learn more about our collaboration project SEISMIC with the University of Surrey!

1. https://pubs.acs.org/doi/10.1021/acs.analchem.3c05677

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