PC-12 Cell Line as a Neuronal Cell Model for Biosensing Applications

Oprea, Daniela; Sanz, Caroline G.; Bârsan, Mădălina M.; Enache, Teodor Adrian

doi:10.3390/bios12070500

Cited by 26 publications

(17 citation statements)

References 114 publications

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“…PC12 cells offer many advantages as they are easily transfectable and have a high yield of CRISPR conversion. Additionally, they display strong responsiveness to differentiation stimuli, such as nerve growth factor (NGF) [ 30 ] or cancer cell supernatants [ 13 , 15 , 19 ], making them an ideal tool for studying neuronal plasticity in vitro [ 13 , 19 , 31 ]. However, most previous studies using this model in cancer biology tracked the stimulation of PC12 cells by soluble factors and vesicles without considering the direct cell-to-cell interactions established at the neuroepithelial interface of cancer cells.…”

Section: Resultsmentioning

confidence: 99%

A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation

Galappaththi

Katz

Howze

et al. 2023

Cancers

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The aggressive nature of certain cancers and their adverse effects on patient outcomes have been linked to cancer innervation, where neurons infiltrate and differentiate within the cancer stroma. Recently we demonstrated how cancer plasticity and TGFβ signaling could promote breast cancer innervation that is associated with increased cancer aggressivity. Despite the promising potential of cancer innervation as a target for anti-cancer therapies, there is currently a significant lack of effective methods to study cancer-induced neuronal differentiation, hindering the development of high-throughput approaches for identifying new targets or pharmacological inhibitors against cancer innervation. To overcome this challenge, we used CRISPR-based endogenous labeling of the neuronal marker β3-tubulin in neuronal precursors to investigate cancer-induced neuronal differentiation in nerve-cancer cocultures and provide a tool that allows for better standardization and reproducibility of studies about cancer-induced innervation. Our approach demonstrated that β3-tubulin gene editing did not affect neuronal behavior and enabled accurate reporting of cancer-induced neuronal differentiation dynamics in high-throughput settings, which makes this approach suitable for screening large cohorts of cells or testing various biological contexts. In a more context-based approach, by combining this method with a cell model of breast cancer epithelial-mesenchymal transition, we revealed the role of cancer cell plasticity in promoting neuronal differentiation, suggesting that cancer innervation represents an underexplored path for epithelial-mesenchymal transition-mediated cancer aggressivity.

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Section: Resultsmentioning

confidence: 99%

A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation

Galappaththi

Katz

Howze

et al. 2023

Cancers

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“…The PC12 cell line is a common in vitro model in neurobiology, which has been used for evaluating the mechanisms involved in Meth neurotoxicity and for assessing potential neuroprotective drugs for use in this context. 25,37,38 Meth affects dopaminergic neurons and causes selective neurodegeneration. Thus, the use of NGF-treated PC12 cells (i.e.…”

Section: Introductionmentioning

confidence: 99%

“…PC12 differentiated cells) represents a valuable method for monitoring Meth-induced neurotoxicity and dopamine release, without having to perform in vivo experiments. [37][38][39] This contributes to a reduction in the numbers of animals used, according to the Three Rs principles. In the current study, a differentiated PC12 cell-based in vitro model was used to investigate the potential neuroprotective properties of TQ against Meth-induced neurotoxicity, by assessing cell viability, apoptosis, the generation of ROS, and the levels of GSH and dopamine.…”

Section: Introductionmentioning

confidence: 99%

Evaluating the Protective Effects of Thymoquinone on Methamphetamine-induced Toxicity in an In Vitro Model Based on Differentiated PC12 Cells

Seyed Aliyan,

Roohbakhsh,

Jafari Fakhrabad

et al. 2024

Altern Lab Anim

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Methamphetamine (Meth) is a highly addictive stimulant. Its potential neurotoxic effects are mediated through various mechanisms, including oxidative stress and the initiation of the apoptotic process. Thymoquinone (TQ), obtained from Nigella sativa seed oil, has extensive antioxidant and anti-apoptotic properties. This study aimed to investigate the potential protective effects of TQ against Meth-induced toxicity by using an in vitro model based on nerve growth factor-differentiated PC12 cells. Cell differentiation was assessed by detecting the presence of a neuronal marker with flow cytometry. The effects of Meth exposure were evaluated in the in vitro neuronal cell-based model via the determination of cell viability (in an MTT assay) and apoptosis (by annexin/propidium iodide staining). The generation of reactive oxygen species (ROS), as well as the levels of glutathione (GSH) and dopamine, were also determined. The model was used to determine the protective effects of 0.5, 1 and 2 μM TQ against Meth-induced toxicity (at 1 mM). The results showed that TQ reduced Meth-induced neurotoxicity, possibly through the inhibition of ROS generation and apoptosis, and by helping to maintain GSH and dopamine levels. Thus, the impact of TQ treatment on Meth-induced neurotoxicity could warrant further investigation.

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“…The PC-12 cell line has been extensively assessed within neuroscience research for its shorter differentiation time and better neurogenic characteristics, as compared to other commercial immortal neuronal cell lines. Enache et al reviewed the applications of different biosensors within PC-12 cell cultures and presented the modern approaches that are employed in neuronal network biosensing [ 4 ].…”

Section: Introductionmentioning

confidence: 99%