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Updated: Mar 15, 2026
An In Vitro Approach to Photodynamic Therapy
Published on: August 17, 2018
5-ALA Fluorescence in Native Pituitary Adenoma Cell Lines: Resection Control and Basis for Photodynamic Therapy
Andrei Nemes1, Thomas Fortmann2, Stephan Poeschke1
1Institute of Neuropathology, University Hospital Muenster, Muenster, Germany.
Objective:
Pituitary adenomas (PA), especially invasive ones, are often not completely resectable. Usage of 5-aminolevulinic acid (5-ALA) for fluorescence guided surgery could improve the rate of total resection and, additionally, open the doors for photodynamic therapy (PDT) in case of unresectable or partially resected PAs. The aim of this study was to investigate the uptake of 5-ALA and the effect of 5-ALA based PDT in cell lines.
Methods:
GH3 and AtT-20 cell lines were incubated with different concentrations of 5-ALA, protoporphyrin IX (PPIX) fluorescence was measured by flow cytometry and fluorescencespectrometry. WST-1 assays were performed to determine the surviving fraction of cells after PDT. PPIX fluorescence intensities and PDT effect of the pituitary adenoma cells were compared to U373MG, a well-known glioblastoma cell line.
Results:
Both cell lines showed a 5-ALA dependent intracellular PPIX fluorescence. Significant differences after 24hrs of incubation were observed in AtT-20 cells in comparison to GH3. Regardless of the incubation or metabolism time, there was a proliferation inhibiting effect after PDT, with no statistical significance.
Conclusion:
Since GH3 cells showed a heterogenous uptake of 5-ALA in the flow cytometry profile, but not constantly high concentrations they might have a 5-ALA efflux mechanism, which still needs to be determined. In the case of AtT-20, the cells might need a longer time for the uptake due to their size or slow metabolism. We showed that the different cell lines have different uptake and metabolism mechanisms, which needs to be further investigated. The general uptake of 5-ALA allows the possibility of resection control and PDT for pituitary adenomas. But, the role of PDT for unresectable pituitary adenomas deserves further investigations.
Insights
5-aminolevulinic acid (5-ALA) shows potential for improving pituitary adenoma resection and photodynamic therapy (PDT). However, cell-specific uptake and metabolism mechanisms require further investigation for optimal therapeutic strategies.
Area of Science:
- Oncology
- Biochemistry
- Cell Biology
Background:
- Pituitary adenomas (PAs), particularly invasive types, present challenges in complete surgical resection.
- 5-aminolevulinic acid (5-ALA) is explored for fluorescence-guided surgery to enhance resection rates.
- 5-ALA also holds potential for photodynamic therapy (PDT) in treating unresectable or residual PA tissues.
Purpose of the Study:
- To investigate the cellular uptake of 5-ALA in pituitary adenoma cell lines.
- To evaluate the efficacy of 5-ALA-based photodynamic therapy (PDT) on these cells.
- To compare 5-ALA uptake and PDT effects between pituitary adenoma and glioblastoma cell lines.
Main Methods:
- Incubation of GH3 and AtT-20 pituitary adenoma cell lines with varying 5-ALA concentrations.
- Measurement of intracellular protoporphyrin IX (PPIX) fluorescence via flow cytometry and spectrometry.
- Assessment of cell viability and proliferation inhibition using WST-1 assays post-PDT, with comparisons to U373MG glioblastoma cells.
Main Results:
- Both pituitary adenoma cell lines demonstrated 5-ALA-dependent intracellular PPIX fluorescence.
- Significant differences in PPIX fluorescence were observed between AtT-20 and GH3 cells after 24 hours of incubation.
- Photodynamic therapy exhibited a proliferation-inhibiting effect across both cell lines, though without statistical significance.
Conclusions:
- Pituitary adenoma cell lines exhibit distinct 5-ALA uptake and metabolism pathways, necessitating further research.
- Potential 5-ALA efflux mechanisms in GH3 cells and slower metabolism in AtT-20 cells warrant detailed investigation.
- While 5-ALA enables potential resection control and PDT for pituitary adenomas, its role in treating unresectable cases requires additional study.
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