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Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy
Miltiadis Fiorentzis1, Periklis Katopodis2, Helen Kalirai3,4
1Department of Ophthalmology, University Hospital Essen, 45147 Essen, Germany.
Abstract:
Three-dimensional (3D) cell cultures represent small avascular tumors in vitro and simulate some of the biological characteristics of solid tumors, enhancing the evaluation of anticancer drug efficacy. Automated image analysis can be used for the assessment of tumor growth and documentation of changes in the size parameters of 3D tumor spheroids following anticancer treatments such as electrochemotherapy. The objective of this article is to assess the effect of various electroporation (EP) conditions (500-750 Volts/cm, 8-20 pulses, 100 µs pulse duration, 5 Hz repetition rate) combined with different bleomycin concentrations (1-2.5 ug/mL) on normal epithelial (HCjE-Gi) and conjunctival melanoma (CRMM1, CRMM2) 3D-cell cultures, through an automated image analysis and a comparison with standard histological assays. A reduction in tumor mass with loss of cell definition was observed after ECT (750 Volts/cm with eight pulses and 500 Volts/cm with 20 pulses) with bleomycin (1 μg/mL and 2.5 μg/mL) in the histological and immunohistochemical analyses of 3D CRMM1 and CRMM2 spheroids, whereas an increase in volume and a decrease in sphericity was documented in the automated image analysis and 3D visualization of both melanoma cell lines. For all other treatment conditions and for the HCjE-Gi cell line, no significant changes to their morphological features were observed. Image analysis with integrated software tools provides an accessible and comprehensive platform for the preliminary selection of homogenous spheroids and for the monitoring of drug efficacy, implementing the traditional screening methods.
Insights
Automated image analysis effectively monitors anticancer drug efficacy in 3D cell cultures. Electrochemotherapy with bleomycin significantly reduced tumor mass in melanoma spheroids, validated by image analysis.
Area of Science:
- Oncology
- Biotechnology
- Medical Imaging
Background:
- Three-dimensional (3D) cell cultures mimic avascular tumors in vitro, aiding anticancer drug efficacy evaluation.
- Automated image analysis offers a method for assessing tumor growth and spheroid changes after treatments like electrochemotherapy.
Purpose of the Study:
- To evaluate the impact of various electroporation (EP) conditions combined with bleomycin on normal epithelial and conjunctival melanoma 3D cell cultures.
- To compare automated image analysis with standard histological assays for assessing treatment effects.
Main Methods:
- Utilized automated image analysis and 3D visualization for monitoring 3D cell cultures (HCjE-Gi, CRMM1, CRMM2).
- Applied various electroporation parameters (500-750 V/cm, 8-20 pulses, 100 µs, 5 Hz) and bleomycin concentrations (1-2.5 µg/mL).
- Conducted histological and immunohistochemical analyses for comparison.
Main Results:
- Electrochemotherapy (ECT) with bleomycin at specific EP conditions (750 V/cm, 8 pulses; 500 V/cm, 20 pulses) and bleomycin concentrations (1-2.5 µg/mL) reduced tumor mass and cell definition in CRMM1 and CRMM2 spheroids.
- Automated image analysis showed increased volume and decreased sphericity in treated melanoma spheroids.
- No significant morphological changes were observed in other treatment conditions or in the HCjE-Gi cell line.
Conclusions:
- Automated image analysis provides a comprehensive platform for preliminary spheroid selection and drug efficacy monitoring.
- This method complements traditional screening approaches in anticancer drug development.
- Specific electrochemotherapy and bleomycin combinations show potential for treating conjunctival melanoma models.
