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Please note that we are not a pharmacy or clinic, so we are unable to see patients and do not offer diagnostic and treatment services for individuals.

In Vitro Cytotoxicity Assay

In vitro cytotoxicity assays are pivotal in preclinical drug development, enabling researchers to assess the safety and therapeutic potential of compounds by measuring their toxic effects on target and non-target cells. As drug discovery shifts toward personalized medicine and complex cell models, Alfa Cytology offers cutting-edge cytotoxicity testing platforms that combine high-throughput automation, 3D organoid systems, and AI-powered data analysis. Our services empower pharmaceutical innovators to optimize candidate selection, reduce attrition rates, and comply with regulatory safety standards.

Introduction to In Vitro Cytotoxicity Assay

In vitro cytotoxicity assays are essential preclinical tools for evaluating the toxic effects of compounds, therapeutic agents, or nanomaterials on cell viability and function. These assays measure dose-dependent toxicity, mechanisms of cell death, and target specificity to guide drug candidate selection. Traditional methods, such as 2D monolayer cultures and endpoint-based MTT assays, are increasingly supplemented by advanced models like 3D tumor spheroids and patient-derived organoids, which better mimic in vivo microenvironments. The rise of personalized medicine and immuno-oncology has expanded their applications, including immune cell-mediated cytotoxicity assessment and nanoparticle safety profiling. Modern approaches integrate multiplexed assays and real-time live-cell imaging for dynamic analysis. Challenges remain in translating 2D data to complex biological systems, driving innovations in AI-driven toxicity prediction and high-throughput screening to enhance scalability and predictive accuracy. As regulatory standards tighten, these assays remain critical for bridging preclinical research and clinical success.

Fig. 1 Cytotoxic functions of CD8 T cells. (Al Moussawy, M., & Abdelsamed, H. A., 2022) Fig. 1 Cytotoxic functions of CD8 T cells. (Al Moussawy, M., & Abdelsamed, H. A., 2022)

Our Services

At Alfa Cytology, we understand that the road to drug discovery is challenging and that ensuring the efficacy and safety of your drug is a crucial step. We are committed to providing you with a comprehensive range of in vitro cytotoxicity assays to help you on your drug development journey.

Co-culture Cell Killing Assays

Alfa Cytology provides specialized co-culture cell killing assays to help clients evaluate the in vitro efficacy of immunotherapeutic drugs. Through the integration of 3D organoid and patient-derived cells (PDC), we support high-throughput screening and dynamic real-time monitoring to comprehensively analyze the tumor-killing ability of effector cells, and provide reliable data close to the clinic for the development of antibody drugs, cell therapies, and drug combination strategies.

T Cell-mediated Killing Assays

Alfa Cytology provides in-depth evaluation of the tumor-specific killing ability of T cells (e.g. CAR-T, CTL). Combined with 3D tumor spheroids, patient-derived organoids and live cell imaging, it quantifies T-cell infiltration, tumor cell apoptosis and cytokine release, providing multi-dimensional data for the optimization of the efficacy of immune cell therapy.

Antibody-dependent Cell-mediated Cytotoxicity (ADCC) Assay

Alfa Cytology focuses on measuring antibody drug-mediated immune cell killing effects. Through multi-color flow analysis with a luciferase reporter gene system, the binding efficiency of the antibody Fc segment to effector cells is dynamically monitored and target cell apoptosis signals are tracked in real time to ensure high sensitivity and reproducibility of data.

Complement-dependent Cytotoxicity (CDC) Assay

Alfa Cytology assesses drug-induced complement-dependent cell killing effects through a standardized experimental process. Using flow cytometry and functional assays, we quantify the formation of the membrane attack complex (MAC) after complement activation and its lysogenic effect on tumor cells, providing key data to support the optimization of antibody drugs.

Common Cytotoxicity Assay Methods

Method	Principle	Advantages	Disadvantages
MTT Assay	Measures mitochondrial activity	Cost-effective, widely validated	Endpoint assay only, no real-time
LDH Release	Detects membrane integrity loss	Quantifies necrosis specifically	Interference from serum components
ATP Detection	Luminescence-based viability	High sensitivity, compatible with 3D	Requires cell lysis
Live Imaging	Isolation by hydrodynamic volume	Dynamic data, visual confirmation	Higher equipment costs

At Alfa Cytology, our end-to-end cytotoxicity testing solutions bridge the gap between early discovery and regulatory submission. By leveraging advanced in vitro models and AI analytics, we help clients de-risk drug candidates, prioritize leads, and accelerate time-to-IND. Contact us today to tailor a cytotoxicity assay strategy that aligns with your drug development goals.

Reference

Al Moussawy, M., & Abdelsamed, H. A. Non-cytotoxic functions of CD8 T cells: "repentance of a serial killer". Frontiers in immunology. 2022, 13: 1001129.

For research use only.