Shan, Feng
(2016)
Investigation of cancer drug penetration in 2D and 3D tumor cell culture models.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
Limited drug penetration into solid tumors may prevent cancer agents from achieving sufficient concentrations to exert their therapeutic effects. However, most pre-clinical in vitro drug discovery studies utilize two dimensional (2D) growth inhibition assays, which do not closely recapitulate either the three dimensional (3D) morphology or cellular microenvironments of solid tumors. In this study, we investigated the penetration of cancer drugs into 2D and 3D head and neck squamous cell carcinoma (HNSCC) cell culture models to determine whether differences in drug penetration might contribute to differences in growth inhibition in the two models. Ellipticine, idarubicin, daunorubicin and doxorubicin were selected as the test drugs because they are fluorescent and a linear relationship exists between fluorescent intensity and drug concentration. Cal33 and FaDu HNSCC cell lines were seeded into normal tissue culture microplates to generate 2D monolayers or Ultra Low Attachment microplates to generate spheroids. We used the Cell Titer Glo® reagent to measure cellular ATP levels as an indicator of cell viability and proliferation. The ImageXpress Micro automated high content platform was used to acquire fluorescent images of 2D and 3D cell cultures exposed to the four fluorescent drugs and the Hoechst DNA stain. We used the multi-wavelength cell scoring image analysis module to quantify the accumulation and distribution of fluorescent drugs in 2D and 3D cell cultures. Although 2D Cal33 cultures were more sensitive than 3D cultures to growth inhibition by all four drugs, both models exhibited similar concentration and time dependent drug accumulation characteristics. In 2D, the distribution of compounds within cells was uniform and independent of the cell numbers seeded into the wells. In Cal33 spheroids however, the compounds exhibited an apparent distribution gradient from high in cells in the outer layers to low in cells in the interior. We speculate that the enhanced cell-cell and cell-ECM contacts in spheroids forms a barrier through which therapeutic agents must penetrate, and that the lower drug concentrations experienced by tumor cells in the interior of spheroids combines with their diminished proliferative capacity to confer the apparent resistance of 3D cultures to cancer drug induced growth inhibition and cell death.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
|
| Date: |
10 August 2016 |
| Date Type: |
Publication |
| Defense Date: |
25 July 2016 |
| Approval Date: |
10 August 2016 |
| Submission Date: |
31 July 2016 |
| Access Restriction: |
No restriction; Release the ETD for access worldwide immediately. |
| Number of Pages: |
117 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Pharmacy > Pharmaceutical Sciences |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
3D tumor cell culture, drug penetration |
| Date Deposited: |
10 Aug 2016 12:23 |
| Last Modified: |
15 Nov 2016 14:35 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/29068 |
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