Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy
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 chang...
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doaj-0e1ba34f213144ffad27f96c912d32d72020-11-25T03:45:16ZengMDPI AGBiomedicines2227-90592020-06-01815815810.3390/biomedicines8060158Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following ElectrochemotherapyMiltiadis Fiorentzis0Periklis Katopodis1Helen Kalirai2Berthold Seitz3Arne Viestenz4Sarah E. Coupland5Department of Ophthalmology, University Hospital Essen, 45147 Essen, GermanyBiosciences, College of Health, Medicine and Life Sciences, Brunel University, London UB8 3PH, UKLiverpool Ocular Oncology Research Group, Department of Molecular and Clinical Cancer Medicine, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3BX, UKDepartment of Ophthalmology, Saarland University Medical Center, 66424 Homburg, GermanyDepartment of Ophthalmology, University Hospital Halle, 06112 Halle, GermanyLiverpool Ocular Oncology Research Group, Department of Molecular and Clinical Cancer Medicine, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3BX, UKThree-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.https://www.mdpi.com/2227-9059/8/6/158conjunctival melanoma3D cell cultureelectroporationelectrochemotherapyspheroid analysisimage analysis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Miltiadis Fiorentzis Periklis Katopodis Helen Kalirai Berthold Seitz Arne Viestenz Sarah E. Coupland |
spellingShingle |
Miltiadis Fiorentzis Periklis Katopodis Helen Kalirai Berthold Seitz Arne Viestenz Sarah E. Coupland Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy Biomedicines conjunctival melanoma 3D cell culture electroporation electrochemotherapy spheroid analysis image analysis |
author_facet |
Miltiadis Fiorentzis Periklis Katopodis Helen Kalirai Berthold Seitz Arne Viestenz Sarah E. Coupland |
author_sort |
Miltiadis Fiorentzis |
title |
Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy |
title_short |
Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy |
title_full |
Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy |
title_fullStr |
Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy |
title_full_unstemmed |
Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy |
title_sort |
image analysis of 3d conjunctival melanoma cell cultures following electrochemotherapy |
publisher |
MDPI AG |
series |
Biomedicines |
issn |
2227-9059 |
publishDate |
2020-06-01 |
description |
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. |
topic |
conjunctival melanoma 3D cell culture electroporation electrochemotherapy spheroid analysis image analysis |
url |
https://www.mdpi.com/2227-9059/8/6/158 |
work_keys_str_mv |
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1724510399047401472 |