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Published on: June 23, 2022
Primary cultures of microglial cells for testing toxicity of anticancer drugs
Mauro Vairano1, Grazia Graziani, Lucio Tentori
1Institute of Pharmacology, Catholic University Medical School, Largo Francesco Vito 1, Rome 00168, Italy.
Abstract:
Toxicity of anticancer agents on normal neural cells during chemotherapy of primary or secondary brain tumors is a clinical problem of increasing relevance and concern. In this perspective, here we used primary cultures of rat cortical microglia as an in vitro paradigm of normal glia to investigate the neurotoxicity of anticancer agents. The effects of two compounds frequently used for treatment of brain tumors, methotrexate (MTX) and temozolomide (TMZ), were compared to those of a known microglial activator, bacterial lipopolysaccharide (LPS); cell viability and metabolism was assessed by the MTS assay. We found that LPS, in the low-intermediate range of concentrations, strongly activates microglia cells, but a highly significant decrease in viability was observed from 100 ng/ml onward. TMZ has no effect at concentrations of clinical interest, whereas MTX significantly increases cell metabolism at 30 microM, a phenomenon possibly reflecting MTX neurotoxicity observed in patients.
Insights
Chemotherapy drugs like methotrexate (MTX) can harm normal brain cells. This study found MTX increased metabolism in microglia cells, suggesting potential neurotoxicity in patients undergoing brain tumor treatment.
Area of Science:
- Neuroscience
- Oncology
- Pharmacology
Background:
- Chemotherapy for brain tumors poses risks to normal neural cells.
- Understanding the neurotoxicity of anticancer agents is crucial for patient safety.
- Microglia are key glial cells in the central nervous system and play roles in neuroinflammation and response to injury.
Purpose of the Study:
- To investigate the in vitro neurotoxicity of anticancer agents on normal glia.
- To compare the effects of methotrexate (MTX) and temozolomide (TMZ) on primary rat cortical microglia.
- To assess the impact of these agents on cell viability and metabolism.
Main Methods:
- Primary cultures of rat cortical microglia were used as an in vitro model.
- Microglia were exposed to methotrexate (MTX), temozolomide (TMZ), and bacterial lipopolysaccharide (LPS).
- Cell viability and metabolism were evaluated using the MTS assay.
Main Results:
- Bacterial lipopolysaccharide (LPS) activated microglia but decreased viability at concentrations ≥100 ng/ml.
- Temozolomide (TMZ) showed no significant effect on microglia at clinically relevant concentrations.
- Methotrexate (MTX) significantly increased microglia cell metabolism at 30 μM, indicating potential neurotoxicity.
Conclusions:
- Methotrexate (MTX) may exhibit neurotoxicity in patients due to its effect on microglia metabolism.
- Temozolomide (TMZ) appears safer for normal neural cells at therapeutic concentrations.
- This study provides in vitro evidence for differential neurotoxic effects of common brain tumor chemotherapeutics.
