Expert Insights on Organoid Research: Best Practices for Growth, Storage, and Monitoring
Organoids offer a powerful platform for translational research, capable of replicating key aspects of human biology that traditional cultures and animal models cannot.1 Their 3D structure mimics the tissue architecture, cellular diversity, and some functional behaviors of real organs, making them increasingly valuable for exploring disease mechanisms and therapeutic responses in a human-relevant context.1
Due to these advantages, organoids have been adopted across a growing range of research areas. However, as their use expands, they place new demands on researchers and the laboratories that support them. Their sensitivity to environmental changes, long culture times, and inherent variability create daily challenges for keeping organoids viable and experiments reliable.
Here, Joe LaPorte, Chief Innovation Officer at PHC Corporation of North America (PHCNA), discusses the key operational challenges researchers face when working with organoids, and the important role of lab infrastructure in reliable organoid research.
Why has the use of organoids increased in recent years?
Over the past decade, advancements in stem cell technologies have allowed us to create organoids that are far more complex and reliable.2 Compared to conventional 2D cultures, they can now give us data that's closer to how real human tissues respond. As a result, organoids are helping researchers cut down on animal testing, run drug screens more efficiently, and avoid expensive failures later in development.2
At the same time, advances in precision medicine and genetic engineering have expanded the scope of organoid applications, attracting an increase in investment from biotechnology and pharmaceutical companies.3 With the future potential to integrate AI into organoid-based research to enable faster drug screening and predictive modeling, it's likely organoids will continue to play a critical role across drug discovery and biomedical research.3
What are the most prominent operational challenges researchers face in maintaining organoid quality and consistency?
Keeping organoids consistent and viable is difficult, primarily because they're living systems that react to even small shifts in their environment. Beyond biological factors, like the stem cell source or media used, day-to-day factors such as how they’re cultured, stored, and monitored can make a big difference to their quality.
One of the key factors is that organoids take time. Their long culture periods mean there's a higher risk of contamination, or variation in conditions that could affect their growth. Every manual step, from media changes to sampling, is another chance for error or variability to creep in.4 And because organoids are so sensitive, even a slight change in culture or storage can alter results.4
Scaling up organoids from small batches to larger volumes can also be quite a challenge. What works in a dish doesn't always translate reliably at scale, especially as there is so much room for variation depending on the conditions.
In short, the operational challenges lie in managing the day-to-day realities of organoid workflows, making sure that systems and equipment function consistently and reliably to keep organoids viable, so your experiments stay reproducible.
How do factors like cold storage and incubation conditions directly impact organoid viability and reproducibility?
Cold storage and incubation conditions can make or break organoid research. First of all, if cold storage temperatures aren't stable, stem cells, media, or reagents may lose viability before cultures even get started, lowering the chances of growing healthy organoids.5 Reliable preservation at the very beginning can be key to getting consistent inputs.5
Once cultures are established, incubation becomes just as important. Variability in temperature, CO2, or oxygen levels can change how organoids grow and differentiate, leading to inconsistent outcomes across batches. And because cultures run for weeks or months, they can be particularly vulnerable to contamination if environments aren’t carefully controlled.
When cryopreserving organoids after growth, reliable ultra-low temperature storage is essential for maintaining viability during freeze–thaw cycles and ensuring organoids can be recovered consistently for future studies.
For researchers, this means that outcomes depend as much on the reliability of storage and incubation infrastructure as it does on the protocols themselves.
Why is real-time monitoring of culture environments important in organoid research?
Due to the long culture times of organoids, it can be easy for small changes in their environment to go unnoticed—and those subtle shifts could affect their viability.
While traditional monitoring approaches rely on periodic sampling or manual checks, which can risk contamination and miss small shifts in conditions, real-time monitoring gives researchers a window into what’s happening as it happens. With continuous data, researchers can identify stress earlier and intervene before cultures fail. It also cuts down on the need for hands-on sampling, which saves valuable staff time and lowers contamination risks.
Additionally, for organoid research, where reproducibility is a major challenge, having consistent, high-resolution data on culture conditions can make it easier to standardize workflows and scale studies further down the line.
What can labs do to help ensure their infrastructure and workflows are reliable for their organoid research?
To ensure lab equipment and workflows are suited to support successful organoid research, labs should:
- Check cold storage reliability: confirm that refrigerators, freezers, and ULTs have strong temperature uniformity and recovery to protect stored materials
- Prioritize incubation performance: choose incubators with precise, stable control of temperature, CO2, and O2, and additional contamination control features such as high heat sterilization, UV, and H2O2 decontamination systems
- Integrate real-time monitoring tools: adopt real-time monitoring systems to reduce manual sampling, lower contamination risks, and generate data that supports confident research standardization
- Utilize proper alarm systems and redundancies: ensure critical equipment is fitted with alarms, remote notifications, and backup systems so issues can be detected and addressed quickly before they compromise cultures or materials
- Ensure systems work together: integrate storage, incubation, monitoring, and support processes into a coordinated workflow to maintain reproducibility and scalability
By focusing on reliability, integration, and proactive safeguards, labs could help reduce variability, protect valuable samples, and generate results they can trust.
References
1 Zhao, Zixuan, et al. "Organoids." Nature Reviews Methods Primers 2.1 (2022): 94.
2 Yang, Siqi, et al. "Organoids: The current status and biomedical applications." MedComm 4.3 (2023): e274.
4 DeStefanis, Rebecca A., et al. "Impact of baseline culture conditions of cancer organoids when determining therapeutic response and tumor heterogeneity." Scientific Reports 12.1 (2022): 5205.
5 Rogulska, Olena, Jarmila Havelkova, and Yuriy Petrenko. "Cryopreservation of organoids." CryoLetters 44.2 (2023): 65-75.