Three-dimensional cell culturing by magnetic levitation for evaluating efficacy/toxicity of photodynamic therapy

Luis G. Sabino; Priscila F. C. Menezes; Vanderlei S. Bagnato; Glauco Souza; Thomas C. Killian; Cristina Kurachi D.D.S.

doi:10.1117/12.2040659

14 March 2014 Three-dimensional cell culturing by magnetic levitation for evaluating efficacy/toxicity of photodynamic therapy

Luis G. Sabino, Priscila F. C. Menezes, Vanderlei S. Bagnato, Glauco Souza, Thomas C. Killian, Cristina Kurachi D.D.S.

Author Affiliations +

Proceedings Volume 8931, Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic Therapy XXIII; 893109 (2014) https://doi.org/10.1117/12.2040659
Event: SPIE BiOS, 2014, San Francisco, California, United States

Abstract

We used three dimensional cell cultures (3D) based on the magnetic levitation method (MLM) to evaluate cytotoxicity of photodynamic therapy (PDT). First, we decorated Hep G2 and MDA-MB-321 cells with NanoShuttle by introducing it in the media and incubated overnight. Next day, we transferred the cells to a 6-well plate and placed a magnetic driver on the top of the plate to start levitation. We monitored the formation of the 3D cell culture by optical microscopy and after four days, we added the photosensitizer Photogem (PG) in the culture media in concentrations of 50, 25, 12.5, 6.25μg/ml. We incubated them for 24 hours, after that we washed the cultures with PBS and added fresh media. Samples were then illuminated for 600s using a 630nm LED-based device, generating light intensities of 30 mW/cm² in a total light fluence of 18 J/cm². Following the illumination, we added fresh media, and 30 hours later, the 3D structures were broken using a pipettor and the cells seeded in 96 well plates, 10⁵ cells per well, with a magnetic drive placed on the bottom of the plate to create cell culture dots. After 24 hours, we used a MTT assay to evaluate PDT cytotoxicity. The PDT effect, evaluated by the half maximal effective concentration (EC₅₀), in MDA-MB-231 cells (EC₅₀ =3.14 μg/ml) is more aggressive compared to the effect of PDT in Hep G2 cells (EC₅₀ = 7.48 μg/ml). It suggests that the cell culture structure and its interaction facilitated the PG uptake and consequently elevated the Photodynamic effect for MDA-MB-231.

Citation Download Citation

Luis G. Sabino, Priscila F. C. Menezes, Vanderlei S. Bagnato, Glauco Souza, Thomas C. Killian, and Cristina Kurachi D.D.S. "Three-dimensional cell culturing by magnetic levitation for evaluating efficacy/toxicity of photodynamic therapy", Proc. SPIE 8931, Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic Therapy XXIII, 893109 (14 March 2014); https://doi.org/10.1117/12.2040659

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available