Cryopreservation: Freezing Time in Cell and Gene

From advanced supply chains and personalized medicines, to interplanetary exploration, cryopreservation could have a profound impact on the future of healthcare and therapeutics.

Advanced therapy developers and researchers are doing their utmost to improve and accelerate the journey of their products from the bench to the bedside. The supporting transport, logistics, and storage specialists tasked with the preservation and delivery of those products are working just as hard to improve their own products and services – and keep pace with a rapidly developing modality. Cryopreservation is seen as a key technology for advanced therapy supply chains, but it’s a technique with many challenges. Here, three experts from the cryopreservation field discuss what is happening now, and what could happen in the future.

Stella Vnook is the CEO of Likarda, a biotech company developing technologies to improve cell therapy delivery, potency, and sustainability. After helping to develop and commercialize a method of coating cells with inert hydrogels to maintain cell viability, Vnook earned a place on our 2024 Power List.

Priya Baraniak is a co-founder and Chief Business Officer of OrganaBio, a biotechnology solutions provider enabling the development and commercialization of cell therapies. Baraniak’s biomedical engineering expertise has culminated in a broad understanding of the cell therapy supply chain, various laboratory techniques, QA/QC measures, GMP lab design & manufacturing, regulatory filings, and technology transfer.

Trevor Smith is the Senior Marketing Manager for Cell & Gene Therapy at Terumo Blood and Cell Technologies. He has expertise in T cell expansion on automated bioreactor systems, process development, GMP lab design, training, and optimized cell collections.

How has the demand for cryopreservation changed with the development of advanced therapies?
 

SV: It has significantly increased. For instance, the global market for preservation equipment was around ($?)2.5 billion in 2014 and is now projected to reach over ($?)8 billion by 2025. This growth has been driven by expansion in cell therapies and regenerative medicine.

We spend so much time and passion launching new therapeutic products, and we want them to be available and accessible. We can scale up manufacturing, but the cost of transportation and logistics are skyrocketing. We need to start solving this before it hits the budget line, which would put us at the mercy of liquid nitrogen transportation.

PB: As we see more and more cell and gene therapies launch, one thing I think we're all very passionate about is their global accessibility and democratization. While we still see a strong preference amongst many cell therapy developers for fresh tissues and cells – especially nascent biotechs – fresh cells pose a real logistics challenge. The transportation and storage of these materials runs against the clock. Cryopreservation enables extended storage and global transport. We’re essentially freezing them in time.

Cryopreserved products have already been embraced by large pharma and established biotech companies. Until we have better processes for lyophilization, or next generation solutions to circumvent cryopreservation, it's going to continue to be the path forward – even to the point of the cryopreservation and transplantation of entire organs.

As we see cell and gene therapies continue to expand in their indications, we're going to continue to see the increased need for cryopreservation technologies, but cost is definitely a major consideration, along with any regulatory hurdles that might exist. The industry, as a whole, needs more standardization and decentralized manufacturing. Cryopreservation does offer some standardization, but the regulations are going to be very important.

TS: There's certainly an increase in demand for cryopreserved products and, as we learn more about these treatments, the more they resemble the very definition of personalized medicines. It's “table stakes” in the operational process, collection, shipping, manufacturing, and reshipping back for reinfusion, but it's no small feat to do all of that. Logistically, it's challenging: aligning schedules for different sites whether in centralized and decentralized manufacturing, but cryopreservation gives you a path forward so these things can get to the patient, where the stakes are high, in time and with the required quality.

Can you elaborate on the advantages of cryopreservation over conventional methods?
 

TS: Conventional methods merely keep the materials fresh or chilled so they remain as close to the native cell and tissue populations as possible. However, they then forego viability risks inherent to the normal freeze-thaw cycle for cells. Cryopreservation stops the clock in manufacturing, easing the pressure of scheduling at the collection site from the patient to better align with the manufacturing availability downstream for more seamless transitions into the manufacturing process. Cryopreservation also enables different dosing strategies, such as multi-dosing, which gives more flexibility for advanced therapy developers to create a product that's more beneficial to patients.

SV: A decade ago, we had very limited cryopreservation tests. Now that cryopreservation is becoming standardized, we expect to optimize it, and testing can go further. We can include cell survival rates, cellular damage tests; we can extend storage duration without compromising efficacy, and we can test at every step how cells are functioning in that environment. A precise and reliable preservation method is critical to ensure therapies that patients receive will have maximum therapeutic benefit.

PB: The IVF field is a major driver of cryopreservation in healthcare and medicine, but beyond fertility, biobanking is still something we do routinely. We have newer options coming to market now, and ideas about first line treatments for patients versus second, third, fourth, fifth line treatments. Today's immunotherapies follow chemotherapy, radiation, monoclonal antibodies or combinations thereof. Many of these patients' bodies are ravaged by the time we are able to go in and perform leukapheresis. Prophylactic apheresis at the point of diagnosis is an emerging idea, and when nothing else has worked, we can try CAR-T therapy using material from the patient from when they were healthier. The hope is that these therapies become first line therapies, rather than last hope chances.

Are there any inhibiting factors that developers might face in accessing or utilizing cryo-based solutions?
 

PB: Bringing automation in will drive down costs and boost standardization. Until then, equipment and equipment costs are definitely an area of limitation. Workforce development is another that needs further attention. Cryobiology is a very specific field; there aren't many cryobiologists out there with the very distinct knowledge of what it takes to cryopreserve cells, tissues, and living materials. You need someone who understands cell biology and the physics of freezing, temperature changes, and nucleation. It's a multidisciplinary field that requires more technical experts to drive it forward.

There are regulatory hurdles, too. Dimethylsulfoxide (DMSO) has been quite ubiquitous and is used routinely in cryopreservation, but we have to make regulators comfortable with the new technologies, materials, and reagents coming to market.

TS: The biggest challenge in adopting automation is that there are so many other areas to focus on during therapeutic development. The final fill and finish step is often deemed a lower priority when compared to expansion or transformation. There is a strong desire to automate and standardize processes, but the timeline for adoptions is often later in clinical trial process development. The primary manufacturing process often gets the most attention.

Priya’s access to expertise point is certainly valid; those in charge of cryopreservation after manufacturing are in a different talent pool to those doing it immediately after collection. How do we train and make it so that both have the same level of expertise, the same consistency, even without the exact same cryopreservation process and starting materials?

SV: There are four factors that are critical. First, the complexity of logistics costs; the infrastructure requirements, temperature control equipment, and backup systems. Second, regulatory and safety concerns. Handling liquid nitrogen comes with risks, as does DMSO. We need to do better as an industry to educate ourselves and our colleagues in the FDA about what we're doing now and where we need to go, and there needs to be more educators. Third, the sustainability factor. How do we create a sustainable solution without excessive energy consumption? And fourth, the environmental impact of the production, transportation, and utilization of liquid nitrogen. Cryopreservation, logistics, and transportation was always thought of as somebody else's headache, until you get to phase III where you start to realize that those costs, implications, and technological hurdles are yours.

A company launching a product has little interest in going back to the beginning to figure out a new way of cryopreserving it. They just want to launch it and make it available. There needs to be an integration of the knowledge accumulated and embedded into earlier stages of the R&D process.

How much optimism is there in the industry when it comes to attracting and developing new talent?
 

TS: The fact that talent is being attracted is a vindication of how compelling the industry is. In 2017, with the first FDA approvals for CAR-T cell therapies, people realized that cell therapies were real, which generated a lot of interest that has compounded in the years since.

However, we need to simplify the training to guide people through the process of pre-programmed protocols. This would lower the barrier to adopting this technology and getting people comfortable with using it. Between the buzz and the excitement around cell and gene therapies, we continue to attract curious individuals, and the advances in science, technology, and automation will become easier to perform and function. Yes, I am very optimistic.

SV: In the past we were looking for people with MDs and PhDs to develop new protocols and systems. What’s more important now is attracting people earlier on; people with bachelor degrees in biochemical engineering or any pharmaceutical process can be motivated by the changes made in the lab. Now these processes are more optimized, we have opportunities for the younger generation earlier in their careers.

They can see how an expansion process translates into a lifesaving medication, as well as how their everyday role translates into saving a life. And if they’re passionate about it, they can continue with their master's and doctorates.

PB: To get the type of workforce that we need for the growth that this industry could see in the next 10 to 25 years, we need to look at programs at the community college level, such as the vocational technical programs we have for other trades. You don't need a full degree; you can learn basic skills on aseptic technique, cryopreservation, and other elements without one. Traditional schooling is not for everyone and we're seeing more and more that a PhD is not paramount. What's more important is attention to detail, hunger and drive, and the need to continue to make science “sexy”.

We've seen a lot of hardship in the industry since 2022, including brutal layoffs and a hard funding landscape. It's incumbent upon leaders in the industry to keep that optimistic outlook and to remain bullish on this, even in the face of adversity. We need to make sure the younger generation isn't turned away. We need to show passion – that inner fire – in our personal missions.

TS: I remember a mentor saying how the next stage will be taking cells out of the body, enhancing it, and putting it back in. I remember another professor saying “that's crazy! It's like science fiction.” But then it happened. The work in this space in the future will certainly be rewarding, and the onus is on us to make people see the connection between manufacturing, collections, and the patient impact. The patient stories are going to become more and more valuable for the field – not just in terms of recruiting, but in keeping the energy and excitement alive.

What do you think cryopreservation is likely to continue to contribute to the advanced therapy sector in the future?
 

SV: It already helps patients in, for example, rural clinics who may not be able to drive to more central locations. Cryo brings the product to them. My mission is to make the treatments available to every person who needs them. Cryo is about removing barriers to access new technologies.

TS: In the near term, I'm hopeful that developers' aspirations for adopting new solutions become a reality. As allogeneic therapies advance into the clinic and beyond, we’ll see more of “off-the-shelf” options become readily available, but scale is going to become the next frontier, and the next problem to solve. Any strides we can make along the way in standardizing how these doses are cryopreserved, stored, and shipped will be critical.

DMSO, a main component of cryo-protected media use, could be phased out. We are looking closely at the emergence of non DMSO-containing media – it's definitely a good space to keep an eye on.

PB: Personalized medicine and the ability to bank cells for future use will become more and more popular. Will we be able to cryopreserve ourselves? Walt Disney has, supposedly! Personally, I wouldn’t want to, but some do!

I can foresee applications in space exploration, and sending cells to space, which is being done by SpaceX and others to discover the effects of microgravity and radiation on different cell types. How might this affect life on the moon or Mars? Maybe someday we will have to leave Earth, or choose to leave Earth. There's a whole new frontier out there in the realm of science fiction, but maybe 50 or 100 years from now, it's conceivable that we might need cryopreservation for things like that.

SV: There's a lot of good we can do with where we are too. Maybe we will reduce the reliance on chemical cryopreservation and move into the era of physical insulators so we can work on better ways of getting blood and organs to those living with conflict. The reality, now, is the instability in the world, and the people facing different challenges. Their priority is to stay alive. If they need blood or any kind of treatment to do that, there are therapies out there. The question is, how do we, as an industry, evolve to be able to deliver?

Similarly, there are parts of the world that need vaccines or other treatments. Expanding our technology box into mechanical, physical insulators and delivering them to where they need to be is a good future to fight for. If we put our heads together, we can solve those challenges.