We recommend avoiding media exchanges when possible.
— Company XOur Technology
Advancing 3D Cell Culture for Drug Discovery, Screening, & Disease Modeling
Explore the scientific rationale and utility provided by current HTMP platforms.
01
The Value
3D cell aggregates provide a more physiologically relevant foundation for drug discovery, screening, and disease modeling.
3D cell aggregates, also known as assembloids, spheroids, microtissues, and 3D cell constructs, are free-floating physiologically relevant 3D cell clusters. In non-adherent microwells, cells self-assemble into an organ-like microarchitecture and reach higher functional maturity than other 2D in vitro culture approaches. Pictured here is a single hepatic spheroid after 4 days in culture, located inside a 600 µm microwell.
Assembloids
Spheroids
Microtissues
3D Constructs
- Improved physiological relevance compared with conventional 2D monolayer culture
- More predictive responses in drug efficacy and toxicity studies
- Broad utility across organ systems for disease modeling and compound screening
02
The Challenge
Current microwell-based platforms lack the ability to adequately change media without disturbing 3D cell culture constructs.
Media exchange is critical for long-term culture and multi-step assays, yet existing platforms consistently fail to retain spheroids during this essential process.
Documented limitations with competitor platforms
Applications that require a media change can be difficult to do without causing spheroid loss.
— Company YThese methods show low throughput and low reproducibility due to the difficulty of changing culture media and the problem of spheroid loss.
— Lee et al., Sci Rep. 2022There is indeed a potential for hypoxic environments at (well) spacings 400 µm and below.
*this refers to how close the center of microwells are to one another. — Wang et al., Advanced Nanobiomed Research, 2022
Cell loss during media exchange leads to compounded viability problems and well to well variability - competitor limitation #1
Inadequate media exchange leads to poor cell health and erroneous results - competitor limitation #2
Microtissue metabolic and oxygen overlap – competitor limitation #3
03
The Solution
HTMP 192
Retention, exchange, and throughput in a single SBS-compatible platform.
HTMP™ 192 combines perfect microwell retention with >85% media exchange in a 192-well format that is SBS / SLAS compatible, enabling physiologically relevant 3D biology at scale.
Through a patent-pending partition wall and microridges, cells remain healthy and undisturbed by fluid flow.
>90%
Microwell Retention
>85%
Media Exchange
192-Wells
SBS / SLAS Compatible
HTMP 192 Solution
Microwell Retention
HTMP 192 maintained spheroid retention across repeated media exchanges compared with competitor platforms.
>85% Media Exchange
Measured exchange performance shows HTMP 192 exceeding the target media exchange threshold.
Cell Health
Representative cell health readout after repeated culture handling and media exchange workflow. Data is normalized to final spheroid number.
Hepatotoxicity Panel
24-hour compound response
Diclofenac (24h)
Troglitazone (24h)
Acetylsalicylic Acid (24h)
300 cells (95% HepG2, 5% LX-2) spheroids were incubated for 6 days prior to a 24-hour compound panel. Viability was assessed following compound incubation using WST-8. Dose-response curves were fit by nonlinear regression (four-parameter logistic).
Application Note
HTMP 192 Brochure
Download the HTMP 192 brochure for platform details, product geometry, and workflow context.
Download BrochureMethodology
For all experiments, 300 cells were seeded per microwell using a 95:5 mixture of HepG2 cells (hepatocyte cell line, Signosis #PC-005) and LX-2 cells (hepatic stellate cell line, Cytion #305039). Co-culture of HepG2 cells with LX-2 cells has been reported to improve baseline 3D aggregate maturity metrics (Pingitore et al., IJMS, 2019). Cells were seeded on day 0, and media exchanges were performed on days 1, 2, 3, and 5 across all platforms. Media was collected on day 6 for albumin quantification by ELISA (Bio-Techne, DY1455). Viability was assessed on day 6, or following a 24-hour compound treatment, by incubating spheroids with 10% WST-8 reagent (APExBIO, K1018) for 3 hours and collecting supernatant for downstream analysis. Spheroids were cultured in phenol red-free, low-glucose (5.5 mM) DMEM (Thermo Fisher, 11054020) supplemented with 10% heat-inactivated fetal bovine serum (Sigma, F4135), 0.5% penicillin-streptomycin (Sigma, P4458), and 15 mM HEPES (Sigma, H0887). All media exchanges were performed using a Vero LTS V-200 pipette (Rainin, 30983910) set to 75 µL/s.
References
Relevant Literature
General Considerations for the Use of New Approach Methodologies in Drug Development Guidance for Industry
U.S. Food and Drug Administration
Guidance for Industry, 2026Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease
Bell & Hendriks et al.
Scientific Reports, 2015High-Fidelity Drug-Induced Liver Injury Screen Using Human Pluripotent Stem Cell–Derived Organoids
Shinozowa & Kimura et al.
Gastroenterology, 2021Prediction of Drug-Induced Hepatotoxicity Using Long-Term Stable Primary Hepatic 3D Spheroid Cultures in Chemically Defined Conditions
Vorrink et al.
Toxicological Sciences, 2018MUST READ
Chemogenomic Screening in a Patient-Derived 3D Fatty Liver Disease Model Reveals the CHRM1-TRPM8 Axis as a Novel Module for Targeted Intervention
Youhanna et al.
Advanced Science, 2024High-throughput automated organoid culture via stem-cell aggregation in microcavity arrays
Brandenburg, Hoehnel, & Kuttler et al.
Nature Biomedical Engineering, 2020The Production of 3D Tumor Spheroids for Cancer Drug Discovery
Sant & Johnston
Drug Discovery Today: Technologies, 2017Reference PDF
Nomenclature
Throughout this website, we may use the terms spheroid and organoid in overlapping contexts. We recognize that these terms carry distinct meanings across the 3D cell culture field and that their usage continues to evolve.
In general, we use spheroid to describe a solid 3D cell aggregate in which cells span from the outer surface toward the inner core. We use organoid to describe a 3D cell aggregate with more tissue-like organization, which may include hollow structures, lumens, or other organ-relevant features.
As the field continues to refine its terminology, our nomenclature may also evolve. We welcome thoughtful feedback from researchers working across spheroid, organoid, and microphysiological system platforms.