A new, fast and semi-automated size determination method (SASDM) for studying multicellular tumor spheroids

Pasha Razifar,Azita Monazzam,Örjan Lindhe,Bengt Långström,Raymond Josephsson,Mats Bergström

doi:10.1186/1475-2867-5-32

Pasha Razifar, Azita Monazzam + Show 4 more

Open Access

https://doi.org/10.1186/1475-2867-5-32

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Abstract

BackgroundConsidering the width and importance of using Multicellular Tumor Spheroids (MTS) in oncology research, size determination of MTSs by an accurate and fast method is essential. In the present study an effective, fast and semi-automated method, SASDM, was developed to determinate the size of MTSs. The method was applied and tested in MTSs of three different cell-lines. Frozen section autoradiography and Hemotoxylin Eosin (H&E) staining was used for further confirmation.ResultsSASDM was shown to be effective, user-friendly, and time efficient, and to be more precise than the traditional methods and it was applicable for MTSs of different cell-lines. Furthermore, the results of image analysis showed high correspondence to the results of autoradiography and staining.ConclusionThe combination of assessment of metabolic condition and image analysis in MTSs provides a good model to evaluate the effect of various anti-cancer treatments.

Highlights

The growth of tumor cells such as Multicellular Tumor Cultures (MTSs) has led to important insights in tumor biology [1]
The total area of the Multicellular Tumor Spheroids (MTS) and total area of necrosis were calculated as the total number of pixels inside the ROIs and was converted into μm2 by using the radius of the reference object
The data represent the mean of 24 MTS

Summary

Introduction

The growth of tumor cells such as Multicellular Tumor Cultures (MTSs) has led to important insights in tumor biology [1]. MTSs represent an intermediary level between monolayer growing cells and solid tumors in animals and humans [2]. MTSs represent quite realistically the three-dimensional growth and organization of solid tumors and simulate the cell-cell interactions and micro environmental conditions found in the tumors [4,5]. Considering the width and importance of using Multicellular Tumor Spheroids (MTS) in oncology research, size determination of MTSs by an accurate and fast method is essential. In the present study an effective, fast and semi-automated method, SASDM, was developed to determinate the size of MTSs. The method was applied and tested in MTSs of three different celllines. Frozen section autoradiography and Hemotoxylin Eosin (H&E) staining was used for further confirmation

Methods

Results

Conclusion