Opening the Black Box: Addressing the Five Key Controversies in Organoids and Organ-on-a-Chip Technologies

On April 10, the FDA announced a plan to "phase out animal testing," stating that it would adopt new technological methods (New Approach Methodologies, NAMs) that are "more effective and more human-relevant" to...Gradually reduce, optimize, or potentially replaceMandatory requirements for animal experiments. NAMs includeAI-based toxicity computational models, cell line computational models, organoids, and organ-on-a-chip.

This groundbreaking regulatory policy will be first applied to the safety evaluation of monoclonal antibodies and will immediately begin to be used for Investigational New Drug (IND) applications, encouraging the inclusion of NAMs data.As early as August 2022, the FDA had approved the world's first new drug IND application based entirely on preclinical data from organ-on-a-chip.

In fact, the implementation of this regulatory policy is a response to the approved bill.——In June 2022, the "2022 Food and Drug Amendment" was passed, marking the first time that organ-on-a-chip and microphysiological systems were included.Independent Non-Clinical Drug Evaluation SystemIncorporated into the bill, organ-on-a-chip and cell models, computer modeling, and animal models are regarded as equally important research methods. In September 2022, the United States passed the FDA Modernization Act 2.0, which eliminated the long-standing requirement in the Federal Food, Drug, and Cosmetic Act that new drugs must undergo mandatory animal testing before human clinical trials. It allows the use of advanced non-animal methods such as cell assays, organ chips, computer modeling, and bioprinting. In December 2024, the U.S. Senate unanimously passed the FDA Modernization Act 3.0, mandating that the FDA update its regulatory guidelines within six months of the bill's passage to promote the implementation of the FDA Modernization Act 2.0.

After the new policy was announced, global social media saw a wave of public opposition, with some experts openly criticizing it, stating that "such an irresponsible move is highly destructive and will lead to chaos in the biopharmaceutical industry." The day after the announcement, model animal CRO companies plummeted, with JOINN Laboratories' A-share hitting a limit-down at the opening bell, while its Hong Kong stock plunged over 18% at one point.

Amidst the torrent, applause and skepticism rise and fall in waves. In response, VCBeat interviewed three rising stars in the organoid/organ-on-a-chip field to discuss thoughts on alternatives to animal testing, standardization, and international collaborations.

Organoids: Adult stem cells or pluripotent cells are cultured in vitro in a three-dimensional system, where cells undergo directed differentiation and self-organize into organ-like structures. These form cell clusters with a certain spatial structure, multiple cell types, and the ability to simulate the functions of real organs. Compared with traditional 2D cell culture models, their 3D cell images are closer to the actual morphology of cells, better simulating complex structures and functions, providing more accurate and reliable experimental models for disease research, drug development, and clinical diagnosis and treatment.

Organoid-on-a-chip and Microphysiological Systems are bioengineering extensions based on the concept of organoids, representing a biomimetic technology rooted in microfluidics and cellular engineering. Essentially, organoid-on-a-chip is a miniaturized cell culture device that leverages a core microfluidic chip platform to achieve reverse biomimicry, enabling high-throughput and precise control of complex physiological environments (such as microenvironments and tissue interfaces). It dynamically cultivates miniature human organs in vitro, replicating key organ functions, and is further applied in dynamic monitoring, high-throughput drug screening, and cellular morphological analysis.

The core of achieving alternatives to animal experiments lies in the cultivation foundation of organoids and organ-on-a-chip — humanized cells. Because they are closer to human physiological characteristics, they can more accurately simulate human responses, thereby reducing experimental deviations caused by cross-species differences.In the FDA's latest release of "The Roadmap to Reducing Animal Testing," it emphasizes that one of the core reasons for reducing preclinical animal experiments isMore than 90% of drugs that appear to be safe and effective in animals fail to gain FDA approval, and some are even fatal to humans.Highlighted the physiological differences between humans and other species.

"Organ chips, during their initial R&D and design phase, are developed as in vitro simulations of the structure and minimal functional units of human organs, with organ function data used to validate their effectiveness. Although there are differences among companies in terms of chip structure and design pathways,Humanization and对标human body are the underlying logic of organoid and organ-on-a-chip development."Ma Chenguang, founder and CEO of Anhui Luohua Biotechnology Co., Ltd., said, 'Many people ask me how the validation effect of organ chips compares to animals. They cannot be compared.'"

Therefore, in human-derived biologics, the immunogenicity issue in animal models may instead cause interference with the real results.In particular, this issue not only may lead to the invalidation of toxicology results but also could hinder the progress of some molecules that should have advanced to clinical trials.

Founder and CEO of Signet Therapeutics, the first innovative targeted drug R&D company in China to combine "organoids + AI," Zhang Haisheng, stated, "Why should the replacement of animal experiments start with monoclonal antibodies? First, as human-derived proteins, monoclonal antibodies have fewer immunogenicity issues in humans, while theoretically, the probability is higher in animal models, highlighting the drawbacks of traditional approaches; second, the systemic toxicity of monoclonal antibody drugs is relatively lower compared to other therapies, and their industrialization level is higher, making pilot risks controllable." Moreover, in terms of monoclonal antibody safety, organoids/organ-on-a-chip can evaluate targeted and non-targeted effects within a controlled human microenvironment.

Another core reason pointed out by the FDA is that developing a monoclonal antibody generally requires the use of 144 non-human primates. As the cost of non-human primates has soared in recent years, adopting more and newer predictive methods can accelerate drug development time and reduce R&D costs.In contrast, organ chips have obvious advantages in high-throughput and large-scale applications, leading to a dual reduction in model costs and time.Miao Chunguang mentioned, "If we equate one chip to one mouse, we can achieve 40 samples on a single chip, which is equivalent to increasing the current throughput by 40 times."

Xie Xin, Co-founder and CEO of Xellar Biosystems, pointed out that high-throughput is embodied in the ability to culture multiple organoids or simulate corresponding organ functions on the same chip or under the same international standard size, thereby efficiently and rapidly conducting early screening of a large number of drugs, which enhances the R&D efficiency of MNCs. Furthermore, the high-quality preclinical database generated after the large-scale application of organ chips, deeply integrated with AI, can build more accurate models for evaluating drug efficacy and safety.

However, as drug development tools, the breakthrough for organoids and organ chips will first occur in the preclinical experimental market where animal models cannot be established or are unstable, such as in certain special tumors, rare diseases, and neurological disorders.

As an example of the world's first new drug approved for IND based on preclinical data from "organ-on-a-chip," in February 2022, Professor Don Ingber (inventor of organ-on-a-chip and member of the three U.S. National Academies) from Harvard University’s Wyss Institute, Dr. Haiqing Bai (co-founder of Xellar Biosystems), and Dr. Longlong Si (currently a researcher at Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, and Director of the Organ-on-a-Chip Joint Laboratory of Xellar Biosystems) jointly led the discovery using lung chips that Azeliragon could significantly block the production of various cytokines causing inflammation. The FDA directly approved its IND application for Phase II clinical trials targeting inflammatory lung diseases.

In June of the same year, Sanofi utilized an organ model developed by Hesperos, which employed induced pluripotent stem cells (iPSC) to differentiate into motor neurons and Schwann cells, obtaining the FDA's second IND based on organ chip data. This also marks the first rare disease chip model that accurately simulates the physiological characteristics of two rare autoimmune demyelinating neurological diseases.

"Organoid chips are not immediately competing with animal models for market share, but rather filling in the gaps, step by step verifying their effectiveness, and achieving a turnaround now and in the future," pointed out Miao Chunguang. Since being recognized as an "independent non-clinical drug evaluation system" in 2022, organ chips and related NAMs...Animal testing can be completely bypassed according to the bill.After the current regulatory details are implemented, the priority of NAMs will be higher than animal testing.—— The "Roadmap" mentions that within 1 to 3 years, by utilizing existing international data, redundant animal experiments will be reduced, shortening the monoclonal antibody animal experiment cycle from 6 months to 3 months. A NAMs database will be established while encouraging companies to submit NAMs data, piloting alternatives to animal experiments, and providing incentives such as priority review. The long-term goal within 5 years is to establish the NAMs database as the default standard, with animal experiments only being used when NAMs cannot address scientific issues.

However, the three practitioners all maintained a wait-and-see attitude towards "completely replacing animal models":In the short term, it is unrealistic for NAMs to capture the market of animal models and experiments. A more likely scenario is that the growth rate of the organoid and organ-on-a-chip markets will increase, with large pharmaceutical companies and CROs expanding their presence, while continuing to explore a comprehensive evaluation system that coexists with traditional methods such as animal models.

Xie Xin said, "Unless humans are used to conduct preclinical experiments, any preclinical model has its own strengths and weaknesses. For instance, organ chips cannot replace animal models in evaluating indicators such as behavioral learning. The best solution is,Apply multiple technical means, cross-validate, and build a comprehensive, cost-effective preclinical drug development system.”

From the perspective of implementation, the FDA's announcement and roadmap this time have moved from directive legislation to a short-term execution plan that includes pilot drugs, incentive measures, and phased programs. This can be seen as the FDA advancing the incorporation of NAMs into routine IND submissions and approvals.

Since 2023, Xellar Biosystems, as the only high-throughput organ chip company, has participated in the OASIS Consortium, the world's most extensive and authoritative collaboration initiated by the FDA, EPA, and more than ten multinational pharmaceutical companies. The consortium focuses on the development of next-generation preclinical drug toxicity prediction tools and the establishment of related standards.

Xie Xin pointed out,"By the end of this year or 2026 at the earliest, more detailed guidelines and consensus standards will be introduced, potentially covering multiple directions such as independent evaluation, safety evaluation, and efficacy evaluation. From the perspective of organ chips, liver single-organ chips applied in toxicology evaluation may become the fastest drug development tool (DDT) to gain FDA recognition and promotion."Recently, a human liver chip has been included in the FDA's Innovative Science and Technology Approaches for New Drugs (ISTAND) pilot program to evaluate its use in predicting drug-induced liver injury (DILI). Data shows that it correctly identified 87% of hepatotoxic drugs that cause liver damage in patients.

It can be seen that,The qualification of Drug Development Tools (DDT) will be the "entry threshold" for the FDA's application of organoid/organ-on-a-chip models.In the specified context of use, DDTs that have obtained qualified certification can be directly applied in corresponding scenarios during drug development and regulatory review (e.g., IND submissions) without requiring the FDA to reconsider or reconfirm their suitability. In other words, for NAMs products, DDT qualification certification serves as a "passport."

For regulators, drug development tools are just a "black box" and do not require high standardization.——For example, the size of organoids or organ-on-a-chip, automated processes, chip uniformity, etc., as well as differences in technical approaches and microfluidic designs among various companies, are not given much attention as long as the output meets certain basic standards and review conclusions.In other words, organoids and organ chips are highly result-oriented and application-oriented.

The formulation of these industry standards is proceeding rapidly. Xie Xin believes that this standard will focus on three directions:

Organoids/Organ-on-a-chip require the cultivation of human-derived cells, where they survive for a certain period and can achieve biochemical indicators and functional validation similar to those of organs/tissues in the human body.

Using currently marketed drugs for control experiments (positive/negative controls) on organoid/organ models, the precision of experimental data can rival or even surpass that of animal models.

Comparison of experimental data with clinical trial results suggests its effectiveness and accuracy, in order to verify its predictive capability for human trial outcomes.

Taking SIGNET's self-developed cardiac organoids as an example, Hai Sheng Zhang and his team collected real patient cardiac toxicity data from more than 120 drugs approved by the FDA or having completed Phase II clinical trials. By comparing preclinical toxicity results with Phase II clinical cardiac toxicity, they discovered limitations in current preclinical cardiac toxicity predictions.The "hERG" gold standard has a very low accuracy rate in reflecting real cardiac toxicity in clinical patients, with only about 40%."The low accuracy cannot be entirely attributed to traditional experimental methods (2D cell models) or animal models; for instance, the hERG index only indicates potassium ion channels."

As a result, the team independently developed a cardiac organoid model with a regularly beating atrial and ventricular structure. The large amount of beating imaging data it generates can be used for AI model training, thereby determining cardiac toxicity by outputting virtual electrocardiogram signals.AI+Cardiac Organoid Drug Toxicity Testing Model Achieves Approximately 85% Accuracy in Blind Drug Testing.

More notably, Zhang Haisheng mentioned that, based on the organoid+AI model, Signet Therapeutics is in contact with leading pharmaceutical companies and may establish in-depth cooperation. Anhui Luohua Biotechnology Co., Ltd.'s full-chain solutions, reagents, and consumables have already reached agreements with overseas pharmaceutical companies. Xellar Biosystems, earlier this year, has...Sanofi, PfizerGlobal pharmaceutical companies have established commercial cooperation.

Xie Xin pointed out, "According to my observation,The top 20 pharmaceutical companies globally have already laid out new tools like organoids/organ-on-a-chip. What they value most is the ability to reduce the cost of large-scale drug screening through high throughput. Secondly, further empowerment via AI+ and other methods enhances the accuracy of model prediction and evaluation.”

In terms of willingness to pay, MNCs certainly have a greater ability to pay for new technologies. However, in the absence of clear regulatory standards, Biotech's investment in paying for such technologies will inevitably be limited, which is also an issue that new tools must face.However, for organoid/organ-on-a-chip R&D enterprises, collaborating with MNCs to gain market recognition and build brand reputation is the top priority.

In 2023,RocheAnnouncement of the Establishment of the Institute of Human Biology (IHB), Focusing on Advancing Research in Human Model Systems Such as Organoids to Accelerate Drug Development. The IHB aims to build a team of 250 people and has invited Professor Hans Clevers, the pioneer of organoids, to join. This makes it the first multinational pharmaceutical giant to heavily invest in this technology.

By the end of 2024,MerckAcquisition of Organoid Development Company HUB Organoids Holding B.V.

Specifically, pharmaceutical companies' applications of organoids/organ-on-a-chip mainly focus on two major directions:

Help pharmaceutical companies screen out pipelines that perform well in safety and efficacy during animal testing but may encounter issues in the clinical stage. This is primarily based on the early simulation of human tissue functions and microphysiological environments using organoids and organ-on-a-chip technologies, reducing the risk of failure after advancing to the clinical stage and potentially saving billions of dollars in R&D costs.

Similarly, "double check" can also be applied in reverse to some pipelines with multiple excellent indicators but only one subpar indicator, further evaluating whether to abandon/advance the clinical pipeline.

Xellar Biosystems combines high-throughput organ chips, cell morphology, multi-omics analysis methods, and computer vision technology to automatically generate 3D high-content biological images of cells, build a 3D cell morphology database, and construct a complex network of interactions between cell morphology, gene clusters, and compound structures for drug repurposing and new drug prediction development.

SIGNET's "Organoid + AI Platform" Deeply Collaborates with XtalPi's "ADME/T Prediction and Screening Based on Deep Neural Networks" in Drug Discovery and Design, Designing and Synthesizing a Batch of Highly Active Candidate Molecules. By Integrating Early AI Design, Optimization, and Preliminary Screening, SIGNET Rapidly Advances Efficacy Evaluation Using Organoid Disease Models and Screens Target Patients Based on Developed Companion Diagnostic Molecular Markers, Achieving Predictable High Response Rates and Addressing Potential Issues in Clinical Stages Before They Arise.

Anhui Luohua Biotechnology Co., Ltd. has independently constructed an organoid biobank containing more than 2,000 cases, over 10 types of common clinical cancers, and more than 30 subtypes of tumor samples and normal tissues. It has also independently developed various chip models such as full-thickness skin chips, lung chips, liver chips, and intestinal chips. Meanwhile, Anhui Luohua Biotechnology Co., Ltd. is developing yet-to-be-conquered pathological model chips and exploring the possibility of connecting multiple single-organ chips in series, such as hyperuricemia involving a three-layer glomerular filtration mechanism.

In terms of going global, countries around the world have set multiple restrictions on the import and export of human-derived cells and stem cells. At the same time, the recent U.S. export restrictions on advanced biotechnology equipment have also significantly impacted cooperation in related upstream life science industries. In response, the three practitioners all hold a cautious attitude, believing thatThe impact of restrictions on cell imports and exports on organoid/organ-on-a-chip collaborations is limited.

Xellar Biosystems has established an R&D center and an FDA/MNC project delivery center in the United States, while in China, it is collaborating with XtalPi to advance the construction of automated laboratory production lines. Cells and data do not cross borders, utilizing local cell sources for R&D activities. The overseas focus of Anhui Luohua Biotechnology Co., Ltd. is more upstream, such as organoid consumables like culture media that are not restricted for import or export. "The chip itself, which does not contain cells, is not restricted for export, so cross-border technical cooperation and solution partnerships remain unaffected." Additionally, outbound business and global exchanges involving cell import/export have sufficient room to operate under relevant domestic approvals.

In terms of collaboration models, they involve multiple application directions such as drug development, cosmetic toxicology, and tumor drug sensitivity testing, which can be summarized as7 Modes of CooperationBased on the differences in development paths, positioning, and commercialization directions, the collaboration models of various organoid/organ-on-a-chip companies also vary.

Licensed products such as single-organ chips, single-disease models composed of multi-organ chips, and the entire technical system solution of AI + models. Collaborators are typically large MNCs, major domestic pharmaceutical companies in China, and Biotechs with a certain market scale. Under this collaboration model, pharmaceutical companies need to have preclinical experimental personnel who can skillfully use the models, thus resulting in relatively higher collaboration and team costs.

From chip models to full-chain products in the entire industry chain, such as culture media and live cell workstations. Compared with customized demands, standardized products have wider application scenarios, including scientific research requiring 3D cell culture and life science solution development.

On the service side, R&D outsourcing and validation outsourcing similar to CRO are important revenue components for organoid/organ-on-a-chip companies. Compared to customized chips or purchasing an entire screening system, the outsourcing model is lower in talent costs, technology patent costs, and short-term time costs.

In-house developed models and related products are offered as a bundled one-time consumables solution to the clients of instrument companies. The underlying logic is that life science instrument R&D companies aim to introduce innovative 3D cell system solutions, but independent R&D is time-consuming and labor-intensive. The OEM approach meets their demand for "core consumables innovation" as a new revenue growth point.

In the clinical stage, tumor organoids directly derived from patient tumor tissues have been proven to be highly consistent with the pathological histological features, molecular characteristics, and drug sensitivity of the patient's tumor. The accuracy of predicting patient treatment outcomes based on their drug sensitivity is relatively high, and they can be used for drug sensitivity testing of most chemotherapy drugs and targeted therapy drugs. In China, this service is mainly operated through third-party laboratories. In recent years, tumor organoid drug sensitivity testing has shown potential for international expansion, with companies exploring the blue ocean markets in Southeast Asia and the Asia-Pacific region.

"TCM enterprises are more receptive and welcoming to organ-on-a-chip than innovative drugs," mentioned Miao Chunguang. Recently, Anhui Luohua Biotechnology Co., Ltd. has established R&D cooperation with several large, time-honored TCM enterprises and multiple TCM research centers, including Qilu Pharmaceutical, Pien Tze Huang, and Ma Yinglong Health Research Institute. "Animal models are the gold standard for Western medicine, partly because of their strong effects, clear mechanisms of action, and the availability of a wider variety of animal models."However, traditional Chinese medicine (TCM) places more emphasis on holism and long-term effects. Coupled with the current limited data requirements for TCM innovative drugs and the adaptability of animal models, there is an urgent need for an alternative model in the research and development process to be applied in toxicology, efficacy, pathology, and other studies.”

Due to the earlier issuance of the "ban" on animal testing in the cosmetics field and more aggressive alternative requirements, the exploration of cosmetic safety assessment using non-animal models has a longer history. The long-term, high-frequency use also demands higher safety verification. Consequently, the exploration of skin organoids and skin-on-a-chip technology began earlier as well.

Taking L'Oréal Group as an example, since 1979, it has been reconstructing human skin models in laboratories to conduct in vitro safety tests as an alternative to animal testing. It also has a research and innovation center in Shanghai, China. According to L'Oréal China, after more than a decade of efforts, the in vitro reconstructed skin has been recognized by various parties as an important alternative testing model. The first multicenter validation of an in vitro skin irritation alternative method based on international validation standards has been carried out in China.

Furthermore, the cosmetics field is exploring efficacy testing using in vitro models such as skin organoids and organ-on-a-chip.Among them, Xellar Biosystems has become a winner of "L'Oréal's 2024 Big Bang Beauty Tech Challenge" with its organ-on-a-chip safety evaluation system in the cosmetics field, and is also collaborating with L'Oréal on related projects.

Thanks to the active promotion by the industry, skin organ chips have become the first model in China to release national standards.In November 2024, the first national standard in China's organ-on-a-chip field, "General Technical Requirements for Skin Chips" (GB/T 44831—2024), drafted by a team led by Gu Zhongze from Southeast University, was officially released. The standard mainly specifies relevant terminology definitions for skin chips, as well as technical requirements for their appearance, cell sources, component performance, and biological performance. It applies to the design, production, and testing of skin chip products based on microfluidic chip platforms.

Overall,Cost-performance ratio, localized market, AI + innovation, mass production (automated production line), and differentiated positioningIs currently the key competitiveness of China-produced organoid/organ-on-a-chip enterprises.

In terms of localization, under the multiple impacts of the China-US trade war, tariff war, and the Illumina风波, innovative enterprises represented by organoids/organ-on-a-chip may becomeThe Next Layout Point for Global Leaders Rooted in the Chinese Market.The logic behind this is,Organoids/Organ-on-a-Chip, as a new tool, is strongly driven by customer needs and application development, with significant localization characteristics.Most customers need to start from scratch, learning new tools and developing entirely new solutions based on requirements, making a localized after-sales service team crucial.

In terms of cost-performance ratio and mass production, several companies interviewed expressed that,Compared with overseas organoids/organ chips, Chinese-produced products have a lower unit price and will be more cost-effective in international cooperation.At the same time, many domestically produced enterprises have built automated production lines, achieving a certain level of standardization and stable mass production.

AI + Innovation and Differentiated Positioning Are Key for Chinese Organoid/Organ Companies to Avoid "Internal Competition."For example, as the first innovative drug R&D company in China with "organoids + AI" as its core technology platform, SIGNET is advancing its independent pipeline development while also engaging in external collaborations for its organoid platform. The phased success of its pipelines continuously validates the scientific and feasibility aspects of the platform. Xellar Biosystems is a platform-based and technology asset-focused company specializing in high-throughput organ-on-a-chip and 3D cell morphology. In the future, it aims to leverage its high-quality database generated by its proprietary platform to promote drug repurposing and new drug discovery. Anhui Luohua Biotechnology Co., Ltd. covers an even more upstream part of the industry chain, providing products, technologies, and services across the entire value chain of organoids/organ-on-a-chip.

To this day, the controversy over the gradual phase-out of mammalian testing for chemical safety continues. The cosmetics industry's transition process offers a clear reference: from breakthroughs in fundamental research to alternative product development, from industrial chain adaptation to adjustments in the regulatory framework. The real key lies not in setting transitional deadlines but in whether emerging alternative technologies can be embraced with an open attitude, driving a paradigm shift from "tool replacement" to "system reconstruction."

Can organoids and organ-on-a-chip tell such stories? We will wait and see.