Beyond the Cage: Enabling Organoid-Based Drug Development
In a landmark shift for biomedical research, the FDA is phasing out its long-standing requirement for animal testing in the development of monoclonal antibodies and other drugs. Announced in an April 10, 2025 statement, this move reflects growing confidence in advanced non-animal models—such as organoids and other in vitro systems—that more accurately predict human responses to new therapies. Enabled by the FDA Modernization Act of 2022, the policy allows the use of alternatives like cell-based assays and computer modeling to support safety and efficacy assessments during early drug development. It also removes the requirement for animal studies in the approval process for biosimilar or interchangeable biological products. This change isn’t just regulatory housekeeping—it’s an inflection point. And for researchers, it’s a green light to explore more ethical, scalable, and human-relevant models. Organoids: The Future, Now Organoids—3D cellular models that mimic the structure and function of human tissues—are leading the charge in this new era. Unlike traditional 2D cultures or animal models, organoids offer researchers a high-fidelity window into disease biology and drug response, all within a controlled, human-relevant system. From oncology to infectious disease research, organoids are rapidly becoming a preferred tool for translational science. However, the promise of organoids hinges on one crucial factor: the quality of the sample. Elevating Organoid Research with Levitation Technology™ At LevitasBio, we recognize that the integrity of your sample can significantly impact the success of an organoid experiment. That’s why we developed the LeviCell® system—to provide researchers with a fast, gentle, and label-free method of cell enrichment that preserves cell viability, phenotype, and function. For organoid researchers, this means: A Match for the Moment As animal testing requirements loosen and organoids gain prominence, LevitasBio is uniquely positioned to facilitate this transition. We’re helping researchers not only meet new regulatory opportunities but push the boundaries of what’s possible in human model systems. Final Thoughts The FDA’s decision is more than a policy update—it’s a call to innovate. As researchers respond with organoid-based strategies, LevitasBio is proud to stand at the intersection of precision, scalability, and biological fidelity. The age of animal-free drug development is here—and it’s time your sample prep caught up. Explore how LeviCell can elevate your organoid workflows: levitasbio.com
The Evolution of Cell Separation and Enrichment: From Centrifugation to Levitation
The ability to isolate, purify, and enrich specific cell populations has transformed biological research, clinical diagnostics, and therapeutic development. Over the centuries, cell separation methods have evolved from rudimentary techniques to sophisticated, high-throughput systems that enable researchers to extract rare and valuable cell populations with unprecedented precision. Early Foundations: The Dawn of Cell Science The journey of cell separation began in the 17th century, when Robert Hooke (1665) and Antonie van Leeuwenhoek (1674) first described cells using early microscopes. While these discoveries laid the groundwork for cell biology, techniques for physically separating cells would not emerge until centuries later. The 20th Century: The Rise of Modern Techniques As biological research advanced, scientists developed methods to separate cells based on physical properties such as size, density, and charge. These early techniques paved the way for modern cell enrichment approaches. The 21st Century: High-Precision and Label-Free Approaches As research needs grew more sophisticated, so did the demand for non-invasive, label-free, and high-throughput cell separation technologies. The Future of Cell Separation As research continues to demand higher purity, efficiency, and scalability, next-generation cell separation methods will likely integrate AI and machine learning. These advancements have the potential to drive innovations in areas such as precision medicine, cell therapy, and regenerative medicine. From the early days of centrifugation to today’s cutting-edge levitation-based separation, cell enrichment technologies have come a long way. At LevitasBio, our mission is to empower researchers by eliminating sample preparation bottlenecks, ensuring every cell matters. By leveraging the power of label-free, gentle Levitation Technology, we enable scientists to obtain the highest quality samples, leading to better data, deeper biological insights, and groundbreaking discoveries. With continued innovation, we are redefining the future of cell enrichment to accelerate scientific breakthroughs across multiple disciplines.
Our North Star for 2025: A Message from Our CEO, Martin Pieprzyk
As we look to the future, I’m excited to share our vision for LevitasBio and the impact we aim to achieve in life science research. Our journey began with a simple yet profound mission: to solve one of the fundamental bottlenecks in life science and empower scientists with the tools they need to uncover biological truth. Today, as new tools and technologies become available and promise to help us advance science, that mission is becoming more critical than ever. At LevitasBio, we deliver life science research solutions that ensure precise, reliable, and reproducible results—enabling the advancement of science towards an accurate understanding of true biology. What does that mean in practice? Let me walk you through where we are today and where we’re headed. A New Era of Life Science Research The life sciences are entering a new era. Scientific breakthroughs are happening at an unprecedented pace, from advancements in single-cell analysis to the growing adoption of organoid and patient-derived models. But with these advancements come new challenges in sample preparation, data integrity, and reproducibility. At LevitasBio, we believe that the foundation of any meaningful scientific discovery starts with the sample itself. Too often, researchers are forced to compromise the quality of the sample during preparation—resulting in compromised data that can delay or even derail critical discoveries. In fact, poor-quality samples are a significant contributor to the over 10,000 scientific publication retractions reported annually1, 2. These retractions often stem from errors linked to compromised data integrity, reproducibility challenges, or flawed methodologies. We believe that many of these issues could be mitigated by fundamentally rethinking how we treat and prepare our most valuable assets—our samples. We aim to ensure that researchers can spend less time on tedious preparation steps and more time focusing on the science that drives innovation. We strive to improve workflows and sample quality by enabling more accurate insights and faster progress. Our Core Values: Precision, Innovation, and Empowerment At the heart of everything we do at LevitasBio are three core values: We are driven by the belief that every researcher should have access to solutions that improve their workflows and enhance their data. The insights gained from their work can revolutionize personalized medicine or help develop novel therapies for some of the most challenging diseases. Looking Ahead: What’s Next for LevitasBio As we look to the future, LevitasBio is focused on expanding our impact across several key areas: New Product Innovations We are committed to developing new technologies that address emerging challenges in sample preparation and cellular analysis. Our pipeline is robust, and we’re excited to share more in the coming months. Collaborations and Partnerships We recognize that scientific progress doesn’t happen in isolation. We are actively building partnerships with leading academic institutions, biotech companies, and research organizations to ensure our solutions are accessible to those who need them most. Empowering Researchers Globally Science is a global endeavor. Our mission is to ensure that researchers worldwide have access to the tools they need to drive discovery. From academic labs to industry leaders, we are dedicated to democratizing access to cutting-edge sample prep solutions. Our Ongoing Commitment to Reproducibility One of the biggest challenges in life science research is reproducibility. Too often, results cannot be replicated due to inconsistencies in sample quality and preparation. This issue doesn’t just slow progress—it erodes trust in science. At LevitasBio, we are committed to solving the reproducibility crisis by providing tools that ensure consistency at every step of the sample preparation process. By delivering solutions that enhance precision and reliability, we help researchers achieve reproducible results, accelerating scientific progress. Closing Thoughts: Driving Discovery Together As I reflect on our journey so far and look ahead to what’s next, I am filled with pride in what we’ve accomplished and excitement for what’s to come. LevitasBio is more than just a company—it’s a community of scientists, innovators, and dreamers passionate about advancing life sciences. Our vision for the future is simple yet profound: to empower researchers to uncover the true biology behind every cell, tissue, and organism. We are committed to providing the tools and support needed to drive discovery and create a lasting impact on human health. Thank you for being part of our journey. Together, we can achieve breakthroughs that were once thought impossible. Let’s make the future of life science research brighter, more precise, and more impactful. Here’s to driving discovery together. Warm regards,Martin PieprzykCEO, LevitasBio Van Noorden, Richard. “More than 10,000 research papers were retracted in 2023 – a new record.” NIH National Library of Medicine, NIH, 2023, More than 10,000 research papers were retracted in 2023 – a new record. Accessed 23 January 2025. Xu, Shaoxiong Brian, and Guangwei Hu. “Worryingly high prevalence of retraction among top-cited researchers.” Nature, Springer Nature, 12 November 2024, https://www.nature.com/articles/d41586-024-03704-8. Accessed 23 January 2025.
New Year’s Resolutions for Researchers: Better Sample Prep, Better Results, and More Discoveries
As the new year begins, researchers worldwide are setting goals to push their science further than ever before. While the typical resolutions around publishing more, writing grants, or networking may already be on your list, we’re here to propose a new approach: focus on your sample preparation workflows. After all, better samples mean better data—and ultimately, better science. Here are some New Year’s resolutions designed specifically for researchers, with a spotlight on new, innovative ways to prepare samples, including organoids and human-subject -derived samples. 1. Embrace Cutting-Edge Sample Prep Technologies Resolve to explore innovative sample preparation technologies to improve my workflow. Traditional sample prep methods often result in cell loss, debris contamination, and compromised viability. In 2025, resolve to try Levitation Technology™ to streamline your workflows. Tools like the LeviCellⓇ system ensure that your samples remain healthy and viable, providing a cleaner and more accurate starting point for downstream analysis. Whether you’re working with organoids, patient-derived xenografts, or other complex samples, there are technologies that can help you achieve: Spend less time troubleshooting and more time generating meaningful results by adopting cutting-edge sample prep tools. 2. Optimize Your Organoid Workflow Refine organoid preparation process to improve consistency and reproducibility. Organoids are becoming a cornerstone of modern research, but their preparation can be challenging. One way to optimize this workflow is by incorporating technologies that improve sample quality at every step. With tools like the LeviCell system, you can: Better organoid prep means more reliable results and faster timelines—a win for any researcher. 3. Reimagine Archival and Cryopreserved Samples Rethink how to approach cryopreserved and archival samples to maximize their value. Many researchers resign themselves to the idea that thawing frozen samples will inevitably lead to a mix of dead and live cells, often accepting suboptimal results. But what if there were ways to salvage more live, viable cells from these precious samples? By optimizing workflows for cryopreserved samples, you can: This approach allows you to unlock the full potential of your archived samples, ensuring that every experiment starts with the best possible material—even from previously frozen samples. It’s a game-changer for researchers who rely on biobanks, human samples, or long-term stored materials. 4. Streamline Your Single-Cell Workflows Focus on improving my single-cell analysis workflows to obtain clearer insights. Single-cell analysis continues to revolutionize research, but it all starts with the quality of your input material. The LeviCell Systems simplify single-cell prep, ensuring that your samples are enriched for live, viable cells before they ever hit your sequencer. With streamlined workflows, you can: This resolution will help you achieve better clarity in your research and avoid costly sequencing errors or oversequencing. 5. Commit to Continuous Learning Stay up to date on the latest advancements in sample prep and cellular analysis. Science is ever-evolving, and staying on top of new technologies and techniques is key to maintaining a competitive edge. Attend webinars, read publications, and participate in vendor shows to learn about the latest innovations in sample preparation and analysis. Some key areas to explore in 2025 include: Ensure your lab is always at the forefront of innovation by committing to continuous learning. 6. Collaborate and Share Insights Continue to collaborate with colleagues and share best practices to improve sample prep across labs. Sharing knowledge is one of the best ways to advance science. In 2025, resolve to collaborate more closely with your peers to share insights into new sample prep methods. Whether through lab meetings, conferences, or online forums, your experiences can help others optimize their workflows and drive more scientific discoveries. Consider hosting a Lunch and Learn or participating in webinars to spread awareness about the latest sample prep solutions. Make 2025 the Year of Better Samples Sample quality is the foundation of meaningful research. By incorporating new technologies and optimizing your workflows, you can ensure your experiments start with the highest-quality samples possible. Here’s to a year of groundbreaking discoveries—starting with better samples! Happy New Year from all of us at LevitasBio! Ready to revolutionize your sample prep workflows? Discover more about how Levitation Technology can help you achieve your research goals in 2025.
Boosting efficiency and productivity with xenograft models
Xenograft models are used extensively in cancer research to simulate the disease’s progression, behavior, and response to treatment. They involve transplanting human cancer cells into immunocompromised mice, providing a controlled environment to study human cancer1. Depending on the research objectives and model type, the transplanted cells may be primary patient-derived tumor or leukemia cells or established cell lines derived from these samples. Patient-derived xenografts (PDX) involve transferring tumor or leukemia cells directly from a patient into an immunodeficient mouse. PDX models retain the original tumor’s histological and genetic features, making them ideal for studying tumor progression2. Cell line-derived xenografts (CDX), on the other hand, use human cancer cell lines cultured in lab conditions and implanted into immunocompromised mice. While CDX models do not fully capture human tumor complexity, their standardization and reproducibility make them valuable for fundamental research and high-throughput screening3. Xenograft models in practice Xenograft models are central to translational medicine attempts, as they help bridge the gap between fundamental in vitro studies and clinical trials. They provide insights into tumor biology, drug efficacy, and resistance mechanisms, supporting the development of more effective targeted therapeutic interventions such as cell therapies. PDX models have proven useful in exploring chemosensitivity and resistance-related targets; studies have highlighted an increase in overall survival and disease-free survival in gallbladder cancer patients treated with PDX-guided chemotherapy4. Moreover, PDX models have helped researchers better understand the molecular characteristics of the cancer landscape in response to targeted therapies5,6. CDX models, on the other hand, are valuable as a primary screening tool in identifying cytotoxic drugs, which can guide further evaluation in more representative models like PDX models7. Challenges in utilizing xenograft models Notwithstanding the importance of xenograft models in cancer research, they come with significant challenges that can hinder their effectiveness. A key hurdle is the initial engraftment of tumor cells into mice (P0 generation). Failure at this stage means losing the sample and the opportunity to explore that specific cancer or subject model. Successful engraftment allows progression to the P1 generation to expand and stabilize the model, but failure here also results in lost potential insights8. The disease must not only engraft but also proliferate in a manner that closely mimics the original human condition. This unpredictable factor can compromise the model’s validity. In CDX models, researchers can modify cells to express specific markers to confirm if the disease is replicating robustly and accurately, but this is not possible in PDX models. It is common for researchers to achieve relatively low and highly variable engraftment efficiencies, limiting disease progression and increasing variability in the animals tested. For example, one study reported an overall engraftment rate of only 27.4% of 113 breast cancer patient samples9. This low efficiency often leads researchers to oversample, which is problematic. In PDX models, the available cells are frequently limited and sometimes of poor quality. While CDX models have more abundant cells, oversampling increases costs significantly due to the expensive maintenance of animal models10. The variability caused by inefficient engraftment complicates data analysis and often necessitates additional replicates, further increasing the number of animals used. Regulatory and ethical considerations also play a critical role, particularly in regions with strict controls like the UK. Organizations such as NC3Rs advocate for reducing animal use, refining procedures to minimize suffering, and replacing animal models when possible11. In general it is best practice that researchers avoid oversampling to minimize animal usage whenever possible. Overcoming xenograft model challenges Achieving high efficiency in engraftment and disease progression, along with consistent disease replication across models, is crucial for reducing turnaround times, simplifying readouts, and minimizing animal use. While engraftment success hinges on a series of uncontrollable factors, such as the sample’s exposure to chemotherapy, the origin of the cells, and the tumor grade9,12,13, evidence has shown that working with highly viable cells can support engraftment, which in turn promotes more rapid disease progression14,15. By thoroughly enriching the sample for the most viable cells and removing dead cells and debris after dissociating tumor cells, toxic effects can be minimized, and engraftment efficiency can be enhanced. Not only can these improvements reduce the quantity of starting material needed per animal, but they can also lead to more effective and rapid disease progression. Ultimately, this approach maximizes the efficiency and ethical use of animals in experiments. Enhancing xenograft efficiency with Levitation Technology To improve xenograft model outcomes, researchers require a cell separation method that can differentiate viable cells from non-viable ones without compromising cell integrity, a challenge often presented by traditional cell separation methods like centrifugation. Levitation TechnologyTM offers a solution; it suspends cells in a paramagnetic solution within a magnetic field, effectively isolating live cells from dead cells and debris while preserving their viability. The LeviCell® system harnesses this technology to support enhanced viability and quality of cells for rodent transplantation, leading to better engraftment efficiency and disease progression. This leads to the generation of more consistent models, allowing researchers to make use of every animal they inject, improving efficiency and productivity while minimizing animal usage11. Dr. Ernesto Diaz-Flores, Associate Professor of Oncology at the University of California San Francisco (UCSF), has successfully integrated Levitation Technology into his team’s workflow for high-risk leukemia CDX models. Previously, the research group’s workflow involved injecting one to two million cells of mixed viability into mice, achieving an 80% engraftment rate and 50% disease progression after five weeks. The LeviCell workflow reduced the number of cells needed to 100,000 at 95% viability, resulting in 100% engraftment and disease progression rates. This not only halved the time to complete engraftment from 10 weeks to five but also enhanced lab productivity and reproducibility of CDX models that accurately reflect disease heterogeneity. References 1. Jin J, Yoshimura K, Sewastjanow-Silva M, Song S, Ajani JA. Challenges and Prospects of Patient-Derived Xenografts for Cancer Research. Cancers. 2023;15(17):4352. doi:10.3390/cancers15174352 2. Hidalgo M, Amant F, Biankin AV, et al. Patient-Derived Xenograft Models: An Emerging Platform for Translational Cancer Research. Cancer Discov. 2014;4(9):998-1013. doi:10.1158/2159-8290.CD-14-0001 3. Souto EP, Dobrolecki LE, Villanueva H, Sikora
Disrupting the Paradigm of Sample Preparation
Sample preparation is a critical step in any cell-based workflow, and its success is pivotal in determining the outcomes of downstream processes such as cell growth for patient-derived xenograft (PDX) models, CRISPR transduction efficiency, single-cell RNA sequencing, proteomics, and functional assays like cytokine release assays (Seahorse) or metabolomics studies (Isoplexis)1,2. At the heart of sample preparation lies cell separation, which needs to be thorough, removing all traces of dead or dying cells and debris, and gentle to preserve cell viability, cell representation, and integrity, all while still maximizing laboratory productivity and research ROI. Researchers continue to encounter challenges with existing cell separation methods, and innovation in the sample preparation space has remained stagnant for over three decades. The current landscape of sample preparation technologies remains dominated by three common methods, however each method presents distinct limitations. As such, it is imperative to consider how innovative approaches to cell separation could address these challenges, thereby improving cell viability, enhancing assay outcomes, and boosting research productivity. Sample preparation technologies Researchers across academia and biotech have long relied on traditional sample preparation technologies, which frequently fail to deliver high-quality outputs. This often results in suboptimal assay outcomes or long exhaustive workflows, particularly with challenging samples. The following sections describe the advantages and limitations of these existing technologies. Centrifugation Centrifugation is a versatile cell separation technique implemented through gradient centrifugation or wash spins3,4. While wash spins are inexpensive and relatively user-friendly, gradient centrifugation requires time and effort for optimization and is highly dependent on the operator’s skill, which introduces variability in results. Cells are susceptible to damage throughout centrifugation, rendering it unsuitable for particularly sensitive cells like primary neurons. Due to their subjective nature, the decanting, aspiration, and resuspension steps further contribute to the inherent variability and enhance the risk and volume of cell loss. Additionally, this method struggles with handling low numbers of cells, making it less suitable for samples with scarce cellular material, such as precious samples from human subjects5. Magnetic beads Magnetic bead-based cell separation is another well-established, cost-effective, and widely accessible method known for its ability to specifically enrich different cell types using targeted magnetic beads6. However, it is limited when working with lower cell numbers and larger cell types (greater than 30 micrometers), and column-based methods are often prone to clogging in the presence of excessive debris or dead cells, resulting in impurities in outputs. The process typically involves multiple steps that require significant hands-on time. Moreover, dead cells must be removed separately using annexin-based methods, which can result in further cell loss7. This can be particularly problematic for cryopreserved samples where annexin-based methods can reduce yields significantly. Flow sorting Flow cytometry-based sorting is a highly sophisticated tool known for its capacity to sort and analyze cells with high specificity8,9. It requires considerable expertise and is often located in core facilities, requiring users to schedule access and limiting availability for frequent use. Flow sorting uses fluorescent labels to identify specific cell types, which can be toxic to some cell types and restrict cell use in particular assays. Moreover, its use is generally limited to smaller cell types (less than 30 micrometers). Flow sorting relies on gating strategies to isolate populations, which can be complicated by large numbers of dead cells or debris, potentially obscuring rare cell types or causing nonspecific reactivity. Additionally, the high pressure used during sorting may not be suitable for sensitive cell types, posing a risk of damaging the cells. Rethinking sample preparation methodologies The limitations of traditional cell separation techniques have significant implications across biomedical research. These limitations require researchers to increase cell numbers to offset the impurities affecting cell counts. Despite these adjustments, cell growth and transduction efficiencies may still be compromised, often resulting in highly variable or inaccurate assay results. Consequently, this can complicate data analysis, render data unclear and unreliable, and may necessitate repeated experiments, thereby consuming valuable time, resources, and funds and causing delays in project timelines. In some cases, these issues can even obstruct the publication of data or hinder the progression of a drug’s development10,11. Innovation is urgently needed to overcome these issues and their significant implications. The ideal cell separation technology would integrate the following key features: A new approach to solving perpetual limitations LevitasBio® has introduced Levitation Technology™ and the LeviCellⓇ platform as a revolutionary solution to address the limitations of traditional cell separation techniques. By levitating live cells in a paramagnetic solution within a magnetic field, they can be separated from dead or dying cells without labeling, minimizing stress and preserving cell integrity. As a result, the LeviCell® systems enhance the viability and quality of target cell populations, which is particularly beneficial for applications such as CRISPR transductions or PDX models, where it supports researchers in achieving improved efficiencies and better model development. This improvement in starting material quality enhances the signal-to-noise ratio and ensures that every sample can directly contribute to meaningful data, reducing the need for extensive data filtering or corrections. This is crucial for precision applications like single-cell RNA sequencing, where each cell’s analysis is essential, or drug dosing assays requiring consistent results to accurately evaluate a drug candidate’s efficacy. The LeviCell system streamlines the cell separation process into three straightforward steps that can be completed in just 20 minutes, significantly shortening timelines, enhancing data validity, and avoiding the waste of resources and research funds. Get in touch today to learn more about how the LeviCell system can support your lab’s research and development needs! References 1. Capelli C, Cuofano C, Pavoni C, et al. Potency assays and biomarkers for cell-based advanced therapy medicinal products. Front Immunol. 2023;14:1186224. doi:10.3389/fimmu.2023.1186224 2. Niepel M, Hafner M, Mills CE, et al. A Multi-center Study on the Reproducibility of Drug-Response Assays in Mammalian Cell Lines. Cell Syst. 2019;9(1):35-48.e5. doi:10.1016/j.cels.2019.06.005 3. Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P. Fractionation of Cells. In: Molecular Biology of the Cell. 4th Edition. Garland Science; 2002. Accessed April 15, 2024. https://www.ncbi.nlm.nih.gov/books/NBK26936/ 4. Harwood R. Cell Separation
Fast-Tracking Cell Therapy Research
Cell-based assays play a central role in the development of safe and effective cell therapies. They serve as methods for quantifying the potency of cell therapies, providing readouts for a broad spectrum of cell therapy characteristics, and are essential throughout the process from initial screening to safety and efficacy testing and dosage optimization1,2. Researchers employ a wide range of advanced cell-based assays in determining cell therapy characteristics, including growth assays, killing assays, metabolic profiling, xenograft models, and molecular profiling3–7. However, despite their critical importance, the application of these assays is fraught with challenges that hinder productivity and impede the progress of cell therapy development, highlighting the need for innovative solutions, like Levitation Technology, to streamline the pathway to therapeutic breakthroughs. Common challenges in cell-based assays: the snowball effect Poor-quality starting material is a primary cause of failure in cell-based assays, characterized by samples containing cells of suboptimal viability, dead or dying cells, and toxic debris. Utilizing low-quality input material undermines research reproducibility and data validity and can trigger a cascade of adverse effects impacting the assay and associated project8. To compensate for the reduced viability and quality of the input material, more cells must be loaded into each well, which becomes particularly problematic when dealing with primary samples or difficult-to-culture cells. Additionally, more biological replicates are required to offset failed attempts and subpar outcomes. Each additional replicate incurs further expense, stretches resources, and demands more hands-on time, leading to lower testing throughput and extended development timelines. Even when experiments are completed under these conditions, the results are often highly variable, and data analysis becomes more complex, further affecting the project’s efficiency and deadlines9. Ultimately, starting with low-quality material expands the scope and cost of projects and diminishes confidence in the outcomes, presenting a significant hurdle to the success of cell therapy projects. Consequently, scientists urgently need effective strategies to overcome these challenges and increase research productivity and outcomes. Improved sample preparation increases productivity Overcoming these significant challenges and enhancing the productivity of cell therapy workflows requires an approach that deals with the root of the problem: sample preparation. A comprehensive approach is needed that addresses several key factors: Historically, the most commonly used cell separation techniques have involved centrifugation, bead, bubble, and fluorescence-activated cell sorting (FACS) approaches16–19. Despite their utility, each of these methods has considerable limitations that exacerbate the previously outlined challenges. For example, bead, bubble, and FACS techniques require cell labeling, introducing additional, time-intensive steps into the protocol and posing the risk of cytotoxicity. Furthermore, these methods often fail to eliminate debris from the samples. Centrifugation, conversely, does not require labeling and is capable of removing debris, yet by nature, it places significant stress on the cells, adversely affecting their viability20. Addressing these limitations is crucial for cell-based work and for in vivo models – enhancing efficiency in cultivating cells for patient-derived xenograft (PDX) models, improving outcomes and timelines in cell-based assays, maximizing research ROI, and ultimately boosting productivity in cell therapy research and development. The answer to increasing productivity – Levitation Technology In a bid to overcome the challenges posed by existing cell separation processes, LevitasBio developed Levitation TechnologyTM, which offers an efficient means of separating cells, substantially improving the outcomes of cell-based assays and boosting productivity. By utilizing a paramagnetic solution within a magnetic field, this technology levitates live cells, distinguishing them from dead or dying cells without the need for labels. Levitation Technology effectively removes debris from samples, yielding data with an improved signal-to-noise ratio compared to other separation methods. In contrast to conventional methods that can impair cell viability and integrity through harsh treatments such as dyes or centrifugation, Levitation Technology offers a gentle alternative. It minimizes cell stress and preserves cell integrity, allowing researchers to enrich for viable cells and major cell-specific subpopulations. Utilizing these high-quality cells in applications such as cell-based assays, CRISPR transductions, or PDX models leads to higher efficiencies, better model growth, and superior assay outcomes. Using high-quality cells or input material opens the door to using fewer cells per reaction. This translates to either testing a wider range of conditions or possibly working with smaller sample sizes. Moreover, the improved starting material quality means that every sample can be used as a data point, and less filtering or data correction is required in downstream analysis. This significantly shortens timelines, enhances data validity, and avoids wasted resources and research funds. This is particularly crucial in applications like single-cell RNA sequencing, where the analysis of each individual cell is critical, or in drug dosing assays, which demand low variability and clean results across time points to accurately determine a drug’s response. By enriching for high viability and removing contaminants, this maximizes every dollar spent on sequencing and yields high-quality results. The LeviCell® platform enables researchers to achieve all these benefits through three simple steps that can be completed in just 20 minutes. Conclusions Cell-based assays are indispensable for developing effective and safe cell therapies, providing crucial insights into therapeutics’ potency, characteristics, and optimal dosages. The challenges that stem from poor-quality cell input can limit the entire development process by necessitating increased resources, time, and financial investment while also compromising the accuracy and reliability of results. Levitation Technology offers a much-needed solution for cell therapy researchers by enhancing cell viability and reducing assay variability. This method leads to improved productivity, efficiency, and reliability. Ultimately, Levitation Technology ensures the generation of high-quality samples, yields better results, and faster turn around in development time which can lead to more effective development of cell therapies. To learn more about how LevitasBio’s LeviCell® platform can help you increase productivity and output in cell-based assays, get in touch with one of our expert team members today! References 1. Capelli C, Cuofano C, Pavoni C, et al. Potency assays and biomarkers for cell-based advanced therapy medicinal products. Front Immunol. 2023;14:1186224. doi:10.3389/fimmu.2023.1186224 2. Salmikangas P, Carlsson B, Klumb C, Reimer T, Thirstrup S. Potency testing of cell and gene therapy products. Front Med. 2023;10:1190016. doi:10.3389/fmed.2023.1190016 3. Eekels JJM,
Exploring the Future of Cell Therapy Research
Cell therapy involves the transfer of viable autologous or allogeneic cells into a patient’s body to replace diseased or damaged cells, modulate cell function, or assist in removing disease-causing or dysfunctional cells1. Cell therapies, encompassing both stem-cell and non-stem-cell therapies, have offered hope to patients and healthcare professionals with their potential to treat diseases once considered incurable2. Despite the promise of cell therapies, researchers and clinicians encounter significant hurdles in the research, development, and manufacturing stages. Here, we will explore recent advancements in cell therapy research, the opportunities and challenges associated with cell-based assays for cell therapy development, and suggest solutions for better outcomes. Recent advancements in cell therapy research Cell therapy has recently witnessed a remarkable transformation owing to technological and methodological advances that have significantly expanded its potential. A noteworthy development has been Chimeric Antigen Receptor (CAR)-T cell therapies, which have shown promise in treating hematological malignancies, offering patients significant improvements in their health and clinical outcomes and the opportunity to go into complete remission from cancer2. However, CAR-T therapy comes with its share of challenges, including potentially adverse effects, limited availability of autologous T cells, and the high cost and time requirements for manufacturing, which have limited its widespread clinical application3. CAR-natural killer (NK) cell therapy has emerged as a potential alternative that is expected to be more well-tolerated and affordable. However, it faces limitations in proliferative capacity, impacting its anti-tumor efficacy4,5. Despite significant advancements, cell therapy remains in its early stages, requiring extensive research efforts to unlock its full potential. To this end, precise, accurate, and efficient cell-based assays are crucial for improving research and manufacturing processes in cell therapy development. Cell-based assays for cell therapy Cell-based assays play a central role in the research and development of cell therapies. Given the complex nature of cell therapy products, assessing a broad spectrum of cell characteristics is essential to accurately predict their clinical effectiveness and safety. In line with this, the FDA requires potency assays to be included in cell therapy testing processes6. Often, measuring cell potency requires several complementary assays to be run in parallel to get a complete picture of the cell therapy’s characteristics and overcome the potential limitations of a single biological assay. Some of the most commonly used cell-based assays relevant for cell potency testing include: The challenge of cell viability Despite their importance for cell therapy research and process development, cell-based assays are not without challenges. The reliability of cell-based assay outcomes is determined by the quality of the starting material, which highlights the critical importance of ensuring only the highest quality cells are input. Ensuring high-quality starting material involves conducting accurate, precise cell counts and viability monitoring15. The presence of dead or dying cells can lead to inaccurate cell counts, meaning that assays may be performed on an insufficient quantity of viable cells. Moreover, dead or dying cells and associated cellular debris and secreted factors can influence an assay’s reactivity and generate misleading signals, compromising the integrity of the results16,17. This can occur through the elevation of background noise, unintentional cellular activation, and transcriptional changes, among other artifacts, which can detract from the assay’s ability to accurately reflect the therapeutic potential and introduce variability in the results. Not only can this lead to samples being wasted and readouts being inaccurate, but it can also lead to researchers drawing the wrong conclusions and cutting a potentially valuable therapy from development due to suspected potency issues when the real problem was with the viability of the test sample. Conversely, it can lead to higher doses being used in patients, which can be problematic in terms of manufacturing and adverse effects. Consequently, cell-based assay manufacturers typically recommend maintaining a minimum viability of 70-90% in the starting material. This ensures unbiased outcomes in high-resolution assays, such as single-cell sequencing and functional metabolic phenotyping, where precision is paramount. However, guaranteeing that cell samples not only attain this viability standard but also do not contain debris or secreted factors can be challenging for researchers and excludes certain techniques such as beads, bubbles, and FACS, which generally do not have the capability to remove debris. The solution: Levitation Technology for enhanced results Levitation TechnologyTM presents an innovative and highly efficient alternative to conventional cell separation methods, significantly enhancing the accuracy and reliability of cell-based assays and the associated data. This technology uses a paramagnetic solution within a magnetic field to levitate viable cells away from dead or dying cells without the need for labels. This method not only simplifies the cell separation process to three straightforward steps, achievable in 20 minutes, but it also ensures high cell viability and yield, crucial for accurate and reliable cell-based assays. Unlike traditional methods involving harsh treatments that impact cell viability and integrity, Levitation Technology offers a label-free, gentle approach to cell enrichment, minimizing cell stress and maintaining integrity. Conclusions and future perspectives Cell therapy represents a transformative treatment that can potentially cure previously incurable diseases. Despite its potential, the field faces research, development, and manufacturing challenges requiring further advancements. Cell-based assays are central to navigating these challenges, offering insights into cell therapies’ potency, efficacy, and safety. Recent technological advancements, like LevitasBioⓇ’s Levitation Technology, further enhance the precision and reliability of these assays by enabling a simple cell enrichment process to obtain the highest quality cells to be used as starting material in these cell-based assays. To learn more about how the LevitasBio LeviCell® platform can support your cell potency assays, get in touch with one of our expert team members today! References 1. El-Kadiry AEH, Rafei M, Shammaa R. Cell Therapy: Types, Regulation, and Clinical Benefits. Front Med. 2021;8:756029. doi:10.3389/fmed.2021.756029 2. Marcuzzi A, Maximova N. Editorial: Advances in stem cell therapy: new applications and innovative therapeutic approaches. Front Med. 2023;10:1225551. doi:10.3389/fmed.2023.1225551 3. Lamers-Kok N, Panella D, Georgoudaki AM, et al. Natural killer cells in clinical development as non-engineered, engineered, and combination therapies. J Hematol OncolJ Hematol Oncol. 2022;15(1):164. doi:10.1186/s13045-022-01382-5 4. Wang X, Yang X, Yuan X, Wang W, Wang Y. Chimeric
Weighing The Risk: The Impact of Inaccuracy in Cell Counting
Stronger Sample Prep Standards Are Essential to Discover Next Gen Cellular Therapies
As a scientist, standardization and appropriately controlling your experiments are vitally important to ensure the questions addressed within those experiments can be answered. We know that too many variables can cause inconclusive results, preventing sample-to-sample comparability. Cell therapy researchers are experiencing variability from donors, patients, tumors, etc. Moreover, the quality of sample material after preparation is often compromised with dead cells and biological debris that can diminish confidence in your data.