Osmolality has long been recognized as a critical quality attribute (CQA) in bioprocessing, encompassing both upstream and downstream workflows. Historically, osmolality measurement has been utilized in protein-based biologics development and manufacturing. It is also crucial for the development and production of cell therapies.

The significance of osmolality testing spans every stage of cell therapy production, from early process development and cell selection to transfection, verification, cryopreservation, storage, and formulation. Osmolality is a quick test that provides versatile information across all phases of cell therapy production about the process efficiency and quality of product.

In this Ask the Expert we spoke with Shweta Nair, Senior Product Manager, Biopharma at Advanced Instruments about the relationship between osmolality and cell therapy with a transcript of our interview below.

What is osmolality, and why is it important to cell therapy?

Osmolality, a measure of solute concentration in a solution, plays a critical role in Cell Therapy. Since cells are highly sensitive to their osmotic environment, maintaining proper osmolality is vital for cell viability.

Monitoring and regulating osmolality throughout the process ensures the cells remain undamaged and avoid stress or lysis (cell bursting). This helps maintain optimal conditions for cell therapy, ultimately impacting its effectiveness.

For example, osmolality is crucial during both cryopreservation and cell culture. During cryopreservation, controlling osmolality helps prevent ice formation, which can be detrimental to cells. In cell culture, proper osmolality supports optimal growth conditions. By ensuring cell integrity and preventing adverse reactions upon administration, osmolality safeguards both the efficacy and safety of cell-based therapies.

How does the osmolality of cell culture media affect cell therapy production?

The osmolality of cell culture media plays a crucial role in cell therapy production, impacting cell viability, growth, and functionality at every stage. Maintaining optimal osmolality is essential to prevent cellular damage: too low can cause cells to swell and burst, while too high can lead to shrinkage and potential death. This balance ensures cells remain viable and functional, supporting their proliferation and differentiation into desired cell types during culture. Proper osmolality also influences metabolic activity and gene expression pathways within cells, crucial for producing therapeutic proteins and maintaining cellular function.

Consistent osmolality is integral to ensuring the quality and safety of cell therapy products. It facilitates stringent quality control measures, minimizing variability in cell characteristics and therapeutic efficacy. Additionally, maintaining precise osmotic conditions enhances process scalability and efficiency, reducing production costs by minimizing cell loss and ensuring batch consistency. Overall, controlling osmolality throughout cell therapy production is fundamental for achieving reliable, effective, and safe cell-based treatments.

From a regulatory perspective, osmolality monitoring is well-established for protein-based therapeutics. Extending this practice to cell therapies ensures comprehensive quality control.

Why is osmolality monitored as a critical process parameter in cell culture?

Osmolality is monitored as a critical process parameter in cell culture for several pivotal reasons. Firstly, it directly influences cell viability by maintaining the osmotic balance crucial for preventing cellular damage from excessive swelling or shrinkage. This ensures cells remain healthy throughout their growth phases. Secondly, optimal osmolality supports efficient cell growth and metabolism, essential for achieving high yields of viable cells in research and therapeutic applications.

Furthermore, monitoring osmolality ensures consistency in experimental conditions, facilitating reliable and reproducible research outcomes. In the context of cell therapy production, precise osmolality control is vital for maintaining the quality and safety of therapeutic cell products. It minimizes variability in cell behavior and characteristics, meeting stringent regulatory standards and ensuring that cell therapies are effective and safe for patients. Overall, osmolality monitoring plays a fundamental role in optimizing cell culture processes, from research settings to clinical applications, to achieve reliable and beneficial outcomes.

How are osmolality and transfection related?

In transfection, osmolality critically optimizes the delivery of genetic material into cells by influencing cell membrane permeability to transfection reagents, thereby enhancing nucleic acid uptake efficiency. Adjusting osmolality not only boosts transfection effectiveness but also preserves cell viability throughout the process, ensuring cells remain healthy and proficient in processing and expressing introduced genetic material post-transfection. This balance is essential for achieving reliable experimental outcomes and high transfection efficiency, crucial for applications such as gene therapy and viral vector production. Researchers often fine-tune osmolality in transfection buffers to optimize delivery while minimizing potential cytotoxic effects.

Research, such as that conducted with Cell and Gene Therapy Catapult in the UK, highlights how increasing salt concentration and osmolality can significantly enhance transfection efficiency. This demonstrates osmolality’s role as a strategic tool in improving transfection processes, underscoring its importance in biomedical research and development. Understanding and optimizing osmolality in transfection protocols are fundamental steps toward ensuring successful gene delivery tailored to the specific requirements of different cell types, thereby advancing the efficacy of transfection techniques across various biomedical applications.

How can osmolality monitoring optimize cell line development and cell banking?

Osmolality monitoring is essential for optimizing both cell line development and cell banking processes. It plays a critical role in selecting robust cell lines by indicating their adaptability to environmental changes, ensuring expected yield and viability throughout development. This monitoring also aids in process development and long-term storage, where maintaining optimal osmolality is crucial for preserving cell integrity and functionality.

In cell banking, osmolality monitoring is particularly vital during sub-banking processes such as freezing and thawing. These procedures can stress cells, potentially compromising their viability and performance. By closely monitoring osmolality, we can optimize these processes to minimize osmotic stress and ensure the robustness of the cell line during storage and subsequent use. This approach not only supports the reliability of research outcomes but also meets regulatory requirements for maintaining consistent quality and safety standards in cell-based applications.

Overall, integrating osmolality monitoring into cell line development and banking practices enhances the resilience and effectiveness of cell cultures, facilitating their successful application in biomedical research and therapeutic innovations.

How can the OsmoTECH^® portfolio of osmometers help cell therapy production?

The OsmoTECH portfolio plays a crucial role in optimizing cell therapy production across multiple stages, spanning from initial cell expansion and production to critical processes like filtration, cryopreservation, freezing, banking, and formulation. Each device within the portfolio is tailored to meet specific throughput requirements at different stages of the production cycle.

For instance, during formulation screening where high-throughput capabilities are essential, our osmometers are equipped to handle large quantities of sample efficiently in industry preferred 96 well plate formats. They are adept at analyzing challenging samples such as cryopreservatives, DMSO, utilizing freezing point depression method endorsed by all pharmacopeias for osmolality testing.

Compliance with regulatory standards is paramount in cell therapy production. Our osmometers are designed to adhere to both non-GMP and GMP guidelines, ensuring robust regulatory compliance and maintaining data integrity throughout the bioprocessing workflow. They are versatile and adaptable to various application needs, delivering the accurate and precise performance expected of analytical instruments in bioprocessing settings.

In summary, the OsmoTECH portfolio of osmometers offers comprehensive support throughout cell therapy production, enhancing efficiency, ensuring quality control, and facilitating regulatory adherence across critical stages of the process.

What are the features of the OsmoTECH^® XT, OsmoTECH^® PRO, and OsmoTECH^® HT, and how do you choose the best fit for your process?

The OsmoTECH^® line from Advanced Instruments offers a range of osmometers (XT, PRO, and HT) designed to cater to specific needs in cell therapy production and bioprocessing. The key factors to consider when choosing the right model are sample throughput, sample complexity, and compliance requirements.

• OsmoTECH^® XT excels in high-precision, single-sample testing. It boasts the widest osmolality range and minimal sample volume requirement, ideal for limited resources and precise measurements. Additionally, its intelligent freezing technology effectively handles even viscous or challenging samples.
• For labs processing multiple samples, the OsmoTECH^® PRO is the perfect choice. This multi-sample micro-osmometer features a 20-position carousel for walk-away testing with small volume requirement.
• OsmoTECH^® HT is the go-to option for high-throughput workflows. It seamlessly integrates with automated systems using a 96-well plate format, significantly boosting efficiency.

OsmoTECH^® instruments go beyond just delivering performance. They prioritize data integrity and GMP compliance, features crucial in our industry. This is achieved through a robust system that includes user management with logins (ensuring clear accountability), audit trails for tracking data changes with timestamps (providing transparency), data security measures like password protection and restricted data transfer (safeguarding against unauthorized access), and electronic signatures for strengthening the audit trail. These features directly support compliance with regulations like 21 CFR Part 11 and EU Annex 11. Additionally, OsmoTECH^® instruments are designed for versatility. They integrate seamlessly into both regulated and non-regulated environments, all while meeting customer needs for effective data management.

For more information, please visit Advanced Instruments

About the Expert

Shweta Nair

Senior Product Manager, Biopharma, Advanced Instruments

Shweta Nair is the senior product manager of the OsmoTECH Bioprocessing Portfolio of Osmometers at Advanced Instruments with over a decade of experience in the biopharma and life sciences sectors. Since 2019, Shweta has led successful development and launch of cutting-edge osmometer products that have redefined accuracy, reliability, data integrity, and performance in osmolality testing for biopharma. Shweta actively engages with thought leaders, industry associations, and regulatory bodies to stay ahead of evolving compliance standards and industry best practices. Prior to Advanced Instruments, Shweta was the global product manager at Millipore Sigma, managing the lab equipment portfolio for biopharmaceutical and research markets.