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