Optimize Cryogenic Storage for Life Sciences

A COMPREHENSIVE GUIDE FOR DECISION MAKING CRYOGENIC STORAGE SOLUTIONS IN LIFE SCIENCES Introduction............................................................................................................................................3 Understanding Cryogenic Storage Options.......................................................................................4 Factors Influencing Storage Decisions...............................................................................................9 On-Site Cryogenic Storage..................................................................................................................11 Automated Cryogenic Storage Systems............................................................................................14 Off-Site Storage Solutions...................................................................................................................17 Choosing the Right Solution................................................................................................................19 Implementing and Managing Cryogenic Storage Systems............................................................21 Case Studies and Success Stories...................................................................................................24 Conclusion............................................................................................................................................28 CONTENTS The National Institutes of Standards in Technology defines cryogenic as temperatures -150° C or lower.¹ These extremely low temperatures influence the rates of water transport and nucleation of cells that maintain sample integrity.² Such low temperatures also ensure the stability of samples during transportation and manufacturing. Numerous studies suggest that even brief instances of warming, arising from temporary exposure to room temperatures, can have detrimental effects on the viability of cellular materials, subsequently influencing patient outcomes.³ ⁴ , 3 In today’s life sciences industry, no company handling biological samples or genetic material can afford to overlook the growing importance of creating a scalable, dependable approach to cryogenic storage. Despite its important role, sample management often takes a backseat due to more immediate concerns, leading many companies to delay planning for its complex and specific requirements. Maintaining sample integrity at ultra-low and cryogenic temperatures entails more than simply employing freezers capable of reaching a specific temperature. It demands a comprehensive plan that takes into account the right type of containers, equipment, and facilities that are equipped with proper ventilation and back-up power supplies. Staff training and labor requirements, as well as maintaining supplies of essential chemicals like liquid nitrogen, must also be part of any plan. These considerations should be factored in from the very beginning to ensure effective decision-making. While most companies recognize the importance of scalable cryogenic storage, they may not fully comprehend the long-term effects that manually accessed storage with limited documentation and traceability could impose on their operations. Additionally, they may overlook potential scalability issues that can arise as they shift from R&D to large-scale clinical trials. By the time they acknowledge the need for space, equipment, and funding, their choices may be limited. That’s why it’s critical to consider all factors that might impact your development and manufacturing process before selecting a cryogenic storage system. This comprehensive guide explores various options, examines the pros and cons of each, and discusses other influential factors that will affect your decision-making. The Science Behind Cryogenics: Why It Matters INTRODUCTION UNDERSTANDING CRYOGENIC 1 STORAGE OPTIONS 5 Understanding Cryogenic Storage Options Manually Accessed or Automatic Cryogenic Storage Systems? MANUALLY ACCESSED CRYOGENIC STORAGE Manual on-site storage, a common choice for small life sciences companies due to its lower startup costs, requires labor-intensive tasks. Personnel are responsible for loading samples, monitoring environmental conditions such as temperature and humidity, recording data, and refilling compartments with liquid nitrogen. Although initially cost-effective, it’s time consuming and prone to human error, risking sample loss or damage. Additionally, manual handling of cryogenic materials poses safety risks. Without adequate protection, employees face exposure to extremely cold temperatures, and potential spills or leaks can create a hazardous environment. From an ergonomic standpoint, manual storage is challenging. The need to use bulky safety equipment, climb ladders, and bend over tanks not only increases the risk of physical injury, but is also time consuming. Locating the correct sample often entails visually searching through multiple racks, possibly exposing other samples to ambient temperatures. This process also increases the risk of samples being misplaced, removed, or damaged. Furthermore, manual systems often lack robust access control, increasing the risk of When it comes to the cryogenic storage of materials, you typically have two choices: house them on-site or establish an agreement with another entity to store them off-site. Both options come with their own set of pros and cons. Wherever you choose, the facility could either be manually operated or equipped with an automatic cryogenic system. Each of those systems also presents its own benefits and drawbacks. unauthorized access or mishandling of samples. The absence of reliable audit trails further complicates matters, making it difficult to accurately track the history of samples. AUTOMATED CRYOGENIC STORAGE Automated cryogenic storage systems leverage software to oversee tasks related to biological sample management. These systems use advanced robotics, automated sample handling, computerized monitoring, and control systems to manage cryopreserved materials. Unlike manually accessed storage freezers, automated cryogenic systems offer heightened precision, significantly minimizing the occurrence of errors due to human oversight. They are also capable of identifying potential bottlenecks and pinpointing opportunities for improved efficiency. The initial investment for these systems can be substantial due to the complex machinery required for automation. However, the long-term benefits – including reduced human error, increased efficiency, and sample integrity – often justify the expense. Particularly when dealing with high-value, critical, or fragile samples, automated systems can provide the best return on investment (ROI). Equipped with sensors and alarms, they maintain stable temperatures even during sample selection and alert users to any irregularities. While the initial investment is higher, the long-term benefits and efficiency gains tend to outweigh these upfront expenses. Space is another key consideration: will an automated system fit in the space allocated? Some automated systems demand larger rooms, additional floor space, special ventilation, utility connections, access aisles, or double-wide doorways. They might also require additional IT management. By taking these factors into account, labs can ensure smooth operation and optimal use of automated cryogenic storage systems. On-Site vs. Off-Site Cryogenic Storage Systems Life sciences companies face a crucial decision on where to store their biological samples: on-site or off-site? Both options offer distinct advantages and disadvantages, necessitating a strategic evaluation based on the company’s specific needs and resources. The decision to either invest in building an in-house facility versus outsourcing, particularly for companies developing novel therapies, is largely influenced by their philosophy and strategy. Many organizations lack the resources and infrastructure to manage all their clinical trial materials, which can range from proteins, cells, and viruses to blood products and bacteria, each with different storage conditions and temperature requirements.⁵ 6 Labor Safety Costs Space Automated Manual Requires personnel to be responsible for loading samples, monitoring temps and humidity levels, recording data, refilling compartments. Poses safety risks such as exposing personnel to extremely cold temperatures and creating a potentially dangerous environment due to the risk of spills or leaks. Low initial startup costs. Low initial startup costs. Leverages software to manage tasks related to biological sample management. This includes advanced robotics, automated sample handling, computerized monitoring, and control systems to manage cryopreserved materials. Heightens precision, significantly minimizing the occurrence of errors resulting from human oversight, detects bottlenecks, and identifies opportunities for increased efficiency. Higher upfront cost that may offer a return on investment only after scaleup or when a large volume of samples are being managed. May require extended height rooms, increased floor space, ventilation systems, utility connections, access aisles, double-wide doorways, and additional IT management. Erica Waller Senior Product Manager Azenta Life Sciences One of the most critical aspects of on-site storage is data management and tracking. Investing in IT infrastructure and sample management tools empowers sample managers to spend less time maintaining their collection and more time focusing on their research. 7 Sue Holland-Crimmin Ph.D., Scientific Consultant Thus, it’s crucial to conduct a comprehensive cost analysis upfront, factoring in hidden expenses like real estate per square foot, facility expenses, automation, labor, and training. This thorough analysis often dictates whether to keep storage in-house or entrust it to external partners. Moreover, certain types of materials might be more suitable for off-site storage due to their specific requirements or the limitations of the organization’s in-house capabilities.⁵ On-Site Storage Offers Convenient Access Biotech and pharma companies often opt for on-site storage of their biological samples, appreciating the convenience and efficiency it provides, especially for highly transacted critical materials. On-site storage is recommended for materials requiring frequent access, assuming that the proper storage conditions, labeling, tracking, and training are in place. In addition, investing in IT sample and inventory management tools can enable scientists to effectively manage and access these materials. However, running an in-house storage facility requires careful planning. Companies must maintain suitable storage conditions and ensure that Compared to small molecules, cells are highly labile. They’re only viable over a narrow range of time and temperature and therefore require justin-time delivery to patients or cryogenic storage to preserve those cells. A lot of organizations haven’t developed that skill in-house. all materials are accurately labeled and tracked. Providing adequate training for staff is also key to managing these materials safely. Investing in IT sample and inventory management tools can provide great value, granting scientists real-time access and control over their inventory, ensuring they have the materials they need, exactly when needed.⁵ Off-Site Storage Provides Process Rigor and Backup Establishing an Optimal Strategy for Biological Sample Storage: A Hybrid Approach Conversely, off-site storage offers advantages like cost savings and improved data integrity. However, a comprehensive cost analysis—considering factors like real estate, facility expenses, automation, manpower, and training—is essential before outsourcing. Materials accessed less frequently or those reserved for disaster recovery, could be better suited for off-site storage with a reliable partner. Numerous organizations specialize in the management, access, and transportation of these materials. Off-site sample management providers often have robust IT infrastructures for material tracking and state-of-the-art automation systems, which are crucial for maintaining and monitoring materials at low temperatures.⁵ To establish an optimal strategy for storing biological samples, biotech and pharma companies may want to consider a hybrid approach, combining both on-site and off-site storage solutions. This method balances convenience and efficiency, especially for frequently transacted critical materials.⁶ In the dynamic world of clinical research, organizations must strategically identify areas where their skill sets and resources can be most effectively utilized to maximize impact. *When choosing an off-site partner, be sure they have systems in place to mitigate any "cons". 8 Sue Holland-Crimmin Ph.D., Scientific Consultant A lot of clinical trials require you to hold specimens for 10 or 15 years. You’re probably not going to hold those within your own facilities. It‘s best to find an appropriate partner to maintain those, and you can access them readily if you need to. On-Site Off-Site Cons Pros • Control • Accessibility • Customization • Upfront costs • Space requirements • Operational responsibilities • Expertise and compliance • Scalability • Predictable costs • Latest technology and Informatics • Limited control • Transportation risks/dependency • Data privacy/management • Disaster backups FACTORS INFLUENCING 2 STORAGE DECISIONS Factors Influencing Storage Decisions 10 Key Takeaway 1 5 2 6 3 7 4 8 What types of samples will you be using? What will your sample volume be like? What kinds of capacity requirements do you have? How easily can you scale up? How can this storage option support your goals of data integrity? How securely can your solution maintain the cryogenic samples and how well can it protect data? Do you have budget for cryogenic storage? How much do you have budgeted for operational expenses, routine maintenance, employees, and software? What kind of consumables and container types are required for the cryogenic samples? Will automation be of interest in the future and if so, what features will support or prevent the use of automation? How reliable is your storage solution? If something goes wrong, how long will you have to fix it, and will immediate human intervention be needed? What kind of maintenance procedures need to be in place? What power-failure backups are in place? What kinds of costs and space considerations do those containers take? What kind of accessibility do you need? How fast do you need to retrieve samples? What regulations and laws do you need to consider with this type of storage? Do you have compliant data accessible? Do you need to keep audit trails that include temperature excursion records? When choosing an automated cryogenic storage system, consider sample volume and capacity, the budget available for upfront and ongoing costs, reliability, accessibility, data integrity, and regulatory compliance. Biological samples, including tissues, cells, DNA, RNA, and proteins, require appropriate cryogenic storage to ensure their integrity and prevent degradation. Numerous factors should be considered when planning how they will be stored: 3 ON-SITE CRYOGENIC STORAGE Considerations for On-Site Storage Advantages of On-Site Cryogenic Storage On-Site Cryogenic Storage Considerations for Emerging Companies On-site cryogenic storage consists of housing all of your cryogenic samples at your company’s facilities. Rather than deploying a software system to regulate all the freezing units, the process is conducted manually. This means that tasks such as temperature monitoring, data collecting, and sample retrieval, among others, are often carried out by personnel. For successful on-site cryogenic storage, you’ll need to consider every variable since you’ll be managing duties internally. Employees, who are the backbone of operations and in high demand for their rare skillset as noted by McKinsey,³ need a facility adequately equipped. Managing the entire process requires you to factor in equipment and maintenance costs, since these won’t be outsourced. Strict adherence to protocols, detailed record-keeping of sample access, transportation locations, and times must be maintained. Training staff on how to avoid transient warming of samples during the selection process is also key. With on-site cryogenic storage, you gain full authority over your samples, letting you set up preferred conditions and specific access guidelines for your team, in line with your facility’s requirements. This ensures any researcher requiring daily access to certain samples can get it. Furthermore, on-site storage offers boundless customization possibilities, enabling you to design your facility according to your precise needs. Small-scale operations have additional considerations to keep in mind. Budget constraints often lead emerging life sciences companies to opt for manual access on-site cryogenic storage, requiring careful allocation for protective equipment, storage containers, and waste management. Despite these financial limitations, safety and maintenance protocols must not be compromised and should be strictly enforced. For smaller companies in product development or preclinical stages, this on-site solution could be adequate, given their typically lesser need for extensive, long-term storage. 12 ● Cryo gloves ● Face shields ● Cryo aprons ● Cryo carriers/carts Types of Cryogenic Equipment and Accessories: ● Dry shippers ● Dewars ● LN2 transfer hoses ● Phase separators Safety training for employees and emergency preparedness plans Regular maintenance of machines Adequate ventilation and insulation Routine cleaning and defrosting Regular calibrations and inspections Proper containerization, filling, and labeling practices Backup systems for power outages Emergency process procedures Handling and Maintenance Protocols Checklist: For more insight on setting up your own cryogenic facility, download our guide “A Practical Guide to Planning a Cryogenic Storage Facility” Resources Key Takeaway Maintaining your own on-site cryogenic storage means you have more control and customization, but you also take on the burden of creating and managing all the protocols yourself. 13 AUTOMATED CRYOGENIC 4 STORAGE SYSTEMS Additional Reading Automated Cryogenic Storage Systems 15 For companies with more extensive storage needs or rigorous digital record-keeping requirements, automated cryogenic storage systems can be game-changing. These systems leverage technology and software to execute numerous tasks, including temperature monitoring, sample retrieval, sample labeling and locating, data recording, and integration, as well as issuing alerts. They provide audit trails that support digital transformation and regulatory compliance. Automated systems also provide an optimal solution for scaling. Their software can detect bottlenecks and identify areas for optimization, thereby fostering rapid growth. Furthermore, automated cryogenic storage systems are available in different sizes, enabling you to select a product that aligns with your company’s size and current scale. For example, Azenta provides automated storage systems that span the spectrum from small enough to fit into a clinical space, such as the CryoArc™ Pico -190°C LN2-Based Automated Storage System, to larger systems such as the CryoArc™ Tera -190°C LN2-Based Automated Storage System, that fulfill larger capacity storage needs. With options designed to cater to collections of various sizes, you can find one that fits your needs the best. Recognizing both your current and future sample storage requirements, along with an awareness of your present and anticipated spatial constraints, is crucial in selecting the optimal system for your needs. An important advantage of automated storage systems is the control of sample exposure to ambient temperatures. Maintaining consistent cryo temperatures during the picking process and avoiding the exposure of innocent samples to transient warming supports greater cell viability.⁴ Read our blog “Cryostorage for Cell Therapies: Addressing Standardizaton Obstacles” to understand how automated cryogenic storage could benefit your company. Why automate? Read our blog to explore the benefits of automated sample storage. Additional Reading Considerations for Types of Compatible Containers When opting for automation, consider the type of containers used in your facility. Automation can be container specific, requiring the use of vials, tubes, bags and racks. Historically, a lack of standardization in cryogenic storage has led to challenges in developing and maintaining consistent handling protocols and thawing processes. For companies considering automation, selecting packaging that adheres to new standards such as those being developed by the International Standards Organization Technical Committee 276 can help ensure a smooth transition and future scalability. Scott Reeves Senior Technical Sales Leader, Azenta Life Sciences Automated cryogenic storage systems revolutionize storage needs and digital record-keeping, enhancing efficiency, scalability, and cell viability, making them indispensable for companies aiming for digital transformation and regulatory compliance. Key Takeaway Despite higher initial costs, automated cryogenic storage systems are beneficial for scalability due to their efficiency and reliable audit trails for regulatory purposes, making them a solid choice for companies in the manufacturing phase of product development. 16 Key Components and Features of an Automated Cryogenic Storage Solution Storage for a variety of types of containers Inventory management software Temperature control, monitoring, and alert systems LIMS connectivity Sample retrieval mechanisms Sample tracking software and integration with other kinds of software OFF-SITE STORAGE 5 SOLUTIONS 18 Security and Risk Mitigation Strategies Advantages of Off-Site Storage Solutions Off-Site Storage Solutions Off-site cryogenic storage is a practical option for organizations without the right skill set or space in-house to manage their own storage. These specialized facilities handle diverse cryogenic samples, accommodating various containers, and product types, often storing samples for multiple companies at once. Despite the benefits of off-site storage, security concerns may arise from not having full control over your products. To alleviate risks associated with offsite cryogenic storage facilities, it is crucial to investigate the following: ● The facility’s emergency preparedness and contingency plans ● Number of on-site personnel and their qualifications ● Reviews and experiences from other companies with this facility ● Types of documentation provided by the facility ● Data gathering methods, analysis processes, and reporting procedures ● Insurance policies and associated costs ● Audit and inspection schedules and requirements ● Data security and encryption measures ● Communication tools for disseminating general repository operations and policies to stakeholders6 Off-site storage facilities can significantly lighten the load of managing intricate, sensitive samples. Companies can entrust them with responsibilities like temperature monitoring, sample retrieval, and data management. These facilities excel in managing daily security details and compliance. They’re not just adept at handling the specifics but also possess robust backup systems, power redundancies, and stringent protocols. Thus, they’re well-prepared for power outages or unexpected situations, ensuring your samples’ safety and integrity through their well-planned contingencies. CHOOSING THE RIGHT 6 SOLUTION Finding the right cryogenic storage solution is not a one-size-fits-all solution. Companies may require a mix of manual access high-efficiency freezers and automated systems, depending on their needs for long-term storage and frequent retrieval. The best choice will depend on various factors such as budget, product development stage, and growth pace. Flexibility will be key to accommodate future expansion and growth. 20 If... And... Then... Your company is in the discovery product development phase. Your company is in the discovery phase. Your primary goal is immediate scalability and maximum efficiency. You’re great at developing products. The company is at an inflection point. The company’s main facility is far away from other facilities. Automated storage may be the right choice. An automated solution can significantly expedite your growth. This technology can pinpoint bottlenecks, highlight areas for efficiency improvements, and oversee inventory and data management. Off-site cryogenic storage is a great option. Facilities that specialize in cryogenic storage offer expertise in compliance and security. They are skilled so you don’t have to be. An automated system is the most effective way to scale quickly if you already have a product in the manufacturing phase and have the capital to invest in a robust system. Having a cryogenic storage facility off-site, closer to your raw materials suppliers, might reduce expenses. However, the risks and potential high costs involved in transferring materials from your central labs to this cryogenic storage facility should not be overlooked. You have the budget and desire to future proof. You are ready to invest in the company to move from the clinical research phase to manufacturing and commercialization/post-market. You are lacking expertise in security or compliance and could use support. You have the resources to fuel explosive growth. Transportation costs are a large factor. Your budget is tight. On-site, manual access cryogenic storage options may be right for you. You’ll need continuous testing and monitoring by a human. Choosing automation-friendly containers to store in can be a good compromise to prepare for future needs. Choosing the Right Solution IMPLEMENTING AND MANAGING 7 CRYOGENIC STORAGE SYSTEMS Plan Logistics: Identify storage needs, product types, scaling speed, supply chain logistics, container needs, and desired storage options before starting. Choose a Location: Opt for a location with minimal exposure to temperature variations, sunlight, and potential hazards. Design the Facility: Ensure sufficient space, ventilation, and insulation to maintain cryogenic temperatures in your facility’s design. Complete Paperwork: Handle insurance, compliance documents, regulatory reviews, and industry protocols early in the process. Acquire Equipment: Invest in top-notch cryogenic storage equipment like freezers, LN2 tanks, cryogenic carriers, and monitoring systems. Implement Software: Install necessary software, including temperature sensors and security systems, and test thoroughly. Establish Routine: Set maintenance schedules, create SOPs, organize cleaning and employee shifts, among other routines. Incorporate Product: Begin with low-risk products testing and ensure system functionality before operating at full capacity. Build Team: Hire personnel for product development, manufacturing, security, etc., as needed. Gather Data: Collect data once product processing begins to identify bottlenecks, improve processes, maintain product quality, and reduce costs. Continuously Improve: Make data-driven decisions and adapt to market changes to enhance product quality and efficiency. When planning and implementing a cryogenic storage system, every detail is important. Your strategy might range from choosing the right storage type to its installation, setup, and regular upkeep. Here’s a possible breakdown of what your plan could include: Plan Logistics Acquire Equipment Build Team Choose Location Implement Software Gather Data Design Facility Establish Routine Continuously Improve Do the Paperwork Incorporate Product 1 5 9 2 6 10 11 3 7 4 8 22 Implementing and Managing Cryogenic Storage Systems Cold Chain Management An article outlining best practices in the Journal of Assisted Reproduction and Genetics recommends that personnel routinely perform spot-check assessments to ensure that LN2 levels are above canister/sample device levels whenever a sample is removed from or placed into a tank.⁷ Companies should also maintain equipment records on-site for the lifespan of equipment like cryogenic storage tanks.⁸ International Society for Biological and Environmental Repositories (ISBER) provides further suggestions for best practices. If your cryogenic facility houses repositories, the organization advises developing clear guidelines that detail the services provided, costs, operating hours, and contact information for stakeholders during regular hours as well as after-hour emergencies.⁶ They also recommend installing O₂ monitoring in areas where LN2 is used.⁶ When it comes to sample tracking and personal protective equipment (PPE), protocols are crucial for ensuring safety and accuracy in labs or research facilities. Precise tracking of samples helps prevent mix-ups and contamination. Every individual involved in the handling of samples must ensure that tracking protocols are followed to the letter. PPE protocols, which protect workers from hazardous materials, must also be taken seriously. Consistent use of proper PPE, in conjunction with appropriate handling and disposal of hazardous materials, can greatly reduce the risk of injury or harm. Cold chain management is an essential part of cryostorage processes. Biological samples are often stored in liquid nitrogen vapor phase (LN2) to maintain temperatures below -135°C, known as the glass transition temperature of water (Tg). It is believed that enzymatic activity stops or is significantly reduced when biosamples are frozen below Tg and remain below Tg until thawed, thus preserving their viability until needed. Currently available commercial LN2 freezers have the capacity to preserve samples at temperatures as low as -190°C. However, once samples are taken out of these storage units, options for their safekeeping and monitoring are limited. It’s crucial that these samples maintain temperatures below their glass transition temperature (Tg) even when they are being handled within the lab environment. The majority of LN2 carriers in the market are bulky and difficult to maneuver, and the sample carriers could potentially present safety hazards. In the absence of automation and software, researchers must carefully time their procedures and have a solid grasp of the warming and cooling rates of cryogenic samples. This knowledge is vital for predicting and managing sample temperatures throughout the entire workflow, enabling researchers to adjust their procedures strategically to prevent samples from crossing the Tg. Samples can cross Tg from -190°C in as few as 45 seconds when exposed to an ambient environment. Insulating the rack and samples during transient exposures helps to slow warming.⁹ Training and education for personnel is always recommended. If you don’t have or need full-time employees for all tasks, service and support plans for preventative maintenance and remote monitoring can be purchased. 23 Monitoring and Maintenance Best Practices ● Check LN2 levels ● Maintain equipment records ● Establish written guidelines ● Install O2 monitoring ● Track samples ● Staff training Best Practice Highlights: CASE STUDIES AND 8 SUCCESS STORIES 25 Automated Cold Chain Infrastructure Boosts Efficiency and Quality in Cryogenic Sample Management In cryogenic storage, cold chain management is the critical component for ensuring long-term sample viability and consistent outcomes, particularly for patients treated with advanced cell and gene therapies. Automation helps support more rigorous, consistent, and repeatable process development. Cold chain management is essential and having an automated system can help manage it more effectively. Despite certain views that automated cold chain infrastructure is an expensive alternative to manual methods, recent research by the Advanced Regenerative Manufacturing Institute (ARMI) BioFab USA proves the ROI value in sample cold chain rigor.⁴ In a workflow involving a manual LN2 dewar, samples from two different shelf locations in a standard rack were monitored during 20 extractions from the freezer over 16 days. The procedure was replicated Figure 1. Experimental Setup. The two experimental conditions: One rack in the Pico and one in an LN, dewar. Each rack had two boxes: a blank box that does not contain cells, and the target box containing the cells to be thawed at the end of the experiment. The blank box was used during each transaction with the target box being removed from the freezer at the beginning and end of the experiment. 0.00 Time (Minutes) 5.00 10.00 15.00 20.00 25.00 2.56 Azenta Dewar 23.77 Target Box Ambient Exposure Time - Cumulative Cryogenic Storage Type Figure 1. Sample Box Placement Within Freezer Racks Experimental Setup and Cell Morphology Results Case Study 1 using Azenta’s CryoArc Pico -190°C LN2-Based Automated Storage System. Following these operations, three vials of induced pluripotent stem cells (iPSCs) from both systems were thawed, expanded, and assessed for cell morphology differences.⁴ The findings suggest that automated storage retains sample quality at least as well as manual methods, while reducing time for sample access, personnel requirements, and PPE usage. Automation significantly minimizes the risk of sample exposure to ambient conditions, as it doesn’t necessitate removing the full rack to access a target location. These results underscore the notable benefits of data accuracy, time savings, and enhanced sample quality through automation, refuting common misconceptions.⁴ Retrieval time for one vial in automated freezer vs manual cryogenic workflow: 2 minutes (automated) vs. 11 minutes (manual). 2VS minutes 11 minutes Get the Poster Figure 3. Moreover, the system safeguards samples in lower shelves from excessive warming by keeping them inside the freezer when accessing boxes in higher shelves.¹¹ Figure 1. The system demonstrated 70% slower warming rates for the first 30 seconds of exposure compared to manual methods, indicating better preservation of sample integrity. The rate of warming in shelf 1 is the highest, but whereas the automated system has relatively linear warming rates for lower shelves, the manual rack has significant differences between different shelves. Figure 2. Sample rate of warming of each shelf from extraction to -120°C (past the glass transition temperature) and the average time to reach -120°C. Another case study from Azenta revealed that automation slows the rate of warming for samples pulled from cryo storage. Liquid nitrogen vapor phase freezers, which keep temperatures below -135°C, are essential for preserving sample viability in biobanking and cell therapy industries. “Innocent samples,” unintended for thawing, can suffer damage due to repeated exposures and thaw/freeze cycles during routine operations. Recent studies compared manual retrieval of cryoboxes to automated handling using Azenta -190°C automated cryogenic storage systems in terms of workflow, time, and warm up rates.¹¹ The study’s findings showed that the automated the system significantly streamlined the process of cryobox removal, compared to manual techniques. Upon rack extraction, the automated system decreased the sample warming rate by 70% within the first half-minute. This system also enabled “innocent samples” to warm across Tg at a 51% slower pace than manual processes. 1 0.00 °C/Minute 5.00 10.00 15.00 20.00 25.00 Manual CryoArc 4 7 9 13 Rate of Warming - First 30 Seconds Shelf Position 1 0.00 °C/Minute 5.00 10.00 15.00 20.00 25.00 Manual CryoArc 4 7 9 13 Rate of Warming from Pull - (~ -180°C) to -120°C Shelf Position Average Time to Warm to -120°C Manual CryoArc 184 Seconds 343 Seconds Case Study 2 26 10 50 80 30 70 100 0 0 1 2 3 4 5 6 7 40 20 60 90 % Confluence Average Confluence Day of Process Control Thaw Averages Manual Thaw Averages Azenta Thaw Averages Read The Full Case Study Sample Warming During Innocent Exposures From an LN2 Freezer: Comparing Temperature, Time & Workflow Using Manual vs. Automated Systems During a 2-minute exposure, the temperatures of all 5 'innocent samples' were compared between manual and automated workflows. The study showed that lower shelf samples warmed less when using the Azenta automated system, especially during the retrieval of a cryobox from the top (shelf 1) and shelf 7. Sample Temperatures Sample Temperatures 27 Time (seconds) Auto Rack Pull #5 120 180 240 Tg at 258s 0 60 -2000 -180.0 -160.0 -140.0 Temperature °C -120.0 -100.0 -80.0 13 10 7 4 1 Time (seconds) Manual Rack Pull #3 120 180 240 Tg at 126s 0 60 -2000 -180.0 -160.0 -140.0 Temperature °C -120.0 -100.0 -80.0 13 10 7 4 1 Time (seconds) Remove Box 7 0 60 120 180 240 -2000 -180.0 -160.0 -140.0 Temperature °C -120.0 -100.0 -80.0 13 10 7 4 1 Time (seconds) Retrieve Box 7 0 60 120 180 240 -2000 -180.0 -160.0 -140.0 Temperature °C -120.0 -100.0 -80.0 13 10 7 4 1 28 Conclusion Whatever you choose, recognize that cryogenic storage is a journey, not a destination. Implementation is a significant milestone, but maintenance will be a constant. The company will scale, markets will change, regulations will be implemented, and new technologies will become available. You’ll need to adapt with ongoing evaluation and adaptation, not just to the outside world, but also to the data you gather to optimize your own facility for continuous improvement. Change is inevitable, so let best practices guide your way. For guidance on making informed decisions about cryogenic storage solutions, discuss your plans with an Azenta expert today. References 1. Dr. Ray Radebaugh. NIST. The MacMillan Encyclopedia Of Chemistry. https://trc.nist.gov/cryogenics/aboutCryogenics.html (2002) 2. Jang, Tae Hoon, Sung Choel Park, Ji Hyun Yang, Jung Yoon Kim, Jae Hong Seok, Ui Seo Park, Chang Won Choi, Sung Ryul Lee, and Jin Han. "Cryopreservation and its clinical applications." Integrative \medicine research 6, no. 1(2017):12-18. 3. Meneghel, J., Kilbride, P. and Morris, G.J. Cryopreservation as a key element in the successful delivery of cell-based therapies—A review. Frontiers in medicine. (2020) 4. Azenta Blog. Considerations for Protecting Cell Viability During Cryopreservation. https://www.azenta.com/resources/benefits-automation-cold-chain-infrastructure (April 2023) 5. Sue Holland-Crimmin, Ph.D. Azenta Resources. The Evolution of Sample Management and Automation in Drug Discovery. https://www.azenta.com/resources/evolution-sample-management-and-automation-drug-discovery (2023) 6. McKinsey & Company. Technology Trends. https://www.mckinsey.com/~/media/mckinsey/business%20functions/mckinsey%20digital/our%20insights/mckinsey%20technology%20trends%20outlook%202023/mckinsey-technology-trends- outlook-2023-v5.pdf. (July 2023) 7. Azenta. Publications & Posters. Benefits of Automation of Cold Chain Infrastructure. https://www.azenta.com/resources/benefits-automation-cold-chain-infrastructure. 8. ISBER. Best Practices: Recommendations for Repositories Fourth Edition. https://cdn.ymaws.com/www.isber.org/resource/resmgr/best_practices_4th_edition/isber_best_practices_recomme.pdf (2018) 9. Azenta. Whitepapers & eBooks. Preserving the Innocents: Biosample Storage at -190ºC. https://www.azenta.com/resources/preserving-innocents-biosample-storage-190-c 10. Azenta. Storage, Automation & Logistics | White Paper. https://web.azenta.com/hubfs/_Current%20Azenta%20Resources/37006-WE-1221-Sample-Warming-During-Innocent-Exposures.pdf (2022) 11. Schiewe, M. C., M. Freeman, J. B. Whitney, M. D. VerMilyea, A. Jones, M. Aguirre, C. Leisinger et al. "Comprehensive assessment of cryogenic storage risk and quality management concerns: best practice guidelines for ART labs." Journal of assisted reproduction and genetics 36 (2019 ): 5-14. ABOUT AZENTA LIFE SCIENCES azenta.com Azenta is a leading provider of life sciences solutions worldwide, enabling impactful breakthroughs and therapies to market faster. Azenta provides a full suite of reliable cold-chain sample management solutions and multiomics services across areas such as drug development, clinical research and advanced cell therapies for the industry's top pharmaceutical, biotech, academic and healthcare institutions globally. Our global team delivers and supports these products and services through our industry-leading brands, including GENEWIZ, FluidX, Ziath, 4titude, Limfinity, FreezerPro, Barkey, and B Medical Systems. Azenta is headquartered in Burlington, MA, with operations in North America, Europe and Asia. For more information, please visit www.azenta.com. © 2024 Azenta US, Inc. All rights reserved. All trademarks are property of Azenta US, Inc. unless otherwise specified. 23005-WE 0824