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PC-12 Cells - The Impact of PC-12 Cells in Neurobiological and Neural Research

PC-12 is an immortalized rat-driven neural cell line used in neurobiology research. It is a valuable research tool for studying the differentiation and proliferation of neural cells. Moreover, researchers utilize these cells to study nerve physiology, cell signalling pathways, and neurotoxicity. Besides, they are also employed in drug testing and development studies.

General information and origin of the PC-12 cell line

Understanding a cell line's general attributes and origin is crucial before commencing work on it. This section will help you learn vital information about the PC12 cell line. Here, you will know: What is the PC12 cell line in pheochromocytoma? How are PC12 cells differentiated? What is the origin of PC12? What is the size of a PC12 cell?

  • PC12 neuronal cells are similar to the primary culture of fetal neurons. They were obtained from pheochromocytoma of the rat adrenal medulla. These cells are of embryonic origin, resembling a mixture of eosinophilic and neuroblastic cells [1].
  • PC12 cells function as catecholamine cells. They synthesize, store, and secrete dopamine and norepinephrine.
  • Pheochromocytoma (PC12) cells have a diameter of 10 to 12 µm.
  • PC-12 cells possess a polygonal-shaped morphology.
  • PC-12 cells have a homogeneous and near-diploid number of chromosomes (n=40).

The greater splanchnic nerve helps with the motility of the foregut and provides sympathetic innervation to the adrenal medulla.

Culturing PC-12 cells

The PC12 cell line neuronal model is widely used in neuroscience research. You should know the following important points to facilitate its easy and effective culturing. This section will include the PC-12 medium, doubling time, seeding density, cell culture protocols, and conditions.

Key Points for Culturing PC-12 Cells

Population Doubling Time:

The PC-12 doubling time is approximately 40 hours.

Adherent or in Suspension:

PC12 neuronal cells grow in small clusters in suspension. They poorly adhere to non-coated surfaces and form patches on collagen.

Seeding Density:

The cell seeding density of the PC 12 cell line is kept at 1 x 104 cells/cm2. For suspension, cells are removed from the substrate by pipetting with a fresh culture medium. To obtain single cells, cells are passed through a 22-gauge needle and poured into new flasks. For cells grown on collagen, wash cells with PBS and incubate in a passaging solution (TrypleExpress) for 10 minutes. Afterward, add fresh media and centrifuge cells. Then, carefully resuspend the pellet and dispense cells into the culture vessel.

Growth Medium:

RPMI 1640 containing 10% fetal bovine serum, 2.1 mM stable Glutamine, and 2.0 g/L NaHCO3 is used to culture the PC 12 cell line. Media is renewed 2 to 3 times per week.

Growth Conditions:

PC-12 cultures are maintained in a 37°C humidified incubator with a 5% CO2 supply.

Storage:

PC-12 cells are stored in the vapor phase of liquid nitrogen or at below -150°C temperature to maintain cell viability for a longer term.

Freezing Process and Medium:

CM-1 or CM-ACF are used for freezing PC12 cells. A slow freezing method is used that allows the temperature to drop by 1°C. This prevents cells from any shock and maintains their viability.

Thawing Process:

Frozen PC-12 cells are thawed in a 37°C water bath. Afterward, cells are added with fresh media and centrifuged. This helps remove freezing media elements. Collected cells are resuspended and dispensed into the flask containing fresh growth medium. Freezing recovery takes almost 24 hours.

Biosafety Level:

Biosafety level 1 laboratory is required to culture the PC12 neuronal cells.

Adherent clusters of PC12 rat pheochromatoma cells observed at 10x and 20x magnifications.

PC-12 Cell Line: Advantages & Limitations

The PC-12 cell line has some distinct features that distinguish it from other neuronal cell lines. This section will delve into the advantages and limitations associated with the use of this cell line in research.

Advantages

  • Easy to Culture: PC-12 cells are robust and relatively easy to culture, making them accessible for various laboratory settings and research purposes.
  • Neuronal Differentiation: They can be induced to differentiate into neuron-like cells, serving as an excellent model for studying neural development, differentiation, and neurogenesis-related processes.
  • Versatility in Research: PC-12 cells are used in a wide range of neurobiological studies, including neurotoxicity, neuroprotection, and the mechanisms underlying neurodegenerative diseases.
  • High Transfection Efficiency: They exhibit high transfection efficiency, facilitating genetic manipulation and making them suitable for studies involving gene expression and function.

Limitations

  • Non-Human Origin: Derived from rat pheochromocytoma, their non-human origin limits their applicability in certain human-specific research areas and may not fully represent human neural physiology.
  • Heterogeneity: Upon differentiation, PC-12 cells can exhibit significant heterogeneity in morphology and function, which may complicate the interpretation of experimental results.
  • Finite Lifespan: While relatively easy to culture, PC-12 cells have a finite lifespan, which may pose challenges for long-term experiments.

PC-12 Cell Line: A Versatile Tool in Neurobiological and Cancer Research

PC12 cell lines, originating from rat pheochromocytoma, serve a pivotal role in neurobiological research, acclaimed for their neuronal characteristic expressions such as neurite outgrowth in response to nerve growth factor (NGF). The versatility of PC12 cells is harnessed extensively in neurobiology for exploring neural differentiation, responses to neurotrophic factors, and neurotoxicity assessments. This unique propensity for differentiation into neuron-like cells upon NGF exposure renders them an indispensable asset for researchers delving into the complexities of the nervous system.

Neuroregeneration and Cell Differentiation

Intriguingly, a study explored the influence of dental pulp stem cells and their derived factors on the differentiation, survival, and expansion of PC12 cells, unveiling promising paths for neuroregeneration [3]. Another investigation spotlighted Osteocalcin, a bone-derived protein known for its calcium-binding capacity, as a promoter of survival, proliferation, and differentiation in PC12 cells [4], highlighting the protein's potential in enhancing neuronal attributes.

Drug Screening and Neuroprotection

The utility of PC12 cells extends to drug screening, particularly for evaluating neuroprotective and neurodegenerative compounds, marking a critical phase in the quest for novel neurodegenerative disease treatments. For instance, research underscored the neuroprotective properties of α-bisabolol in a Parkinson's disease model employing PC12 cells, evidencing α-bisabolol's capability to mitigate amyloidogenesis and apoptosis while also modulating the neurotoxic effects of amyloid beta [5].

Cellular Transformation and Neural Networks

The process of differentiation from a PC-12 cell to a neuron-like entity commences with NGF, a pivotal protein essential for neuronal growth and survival. Exposure of PC-12 cells to NGF activates complex intracellular pathways, laying the groundwork for their transformation. This metamorphosis is a gradual revelation of cellular potential, observable under a microscope as the cells extend neurites and forge connections akin to neuronal networks.

Morphological Changes and Neuronal Phenotype

Differentiation imparts significant morphological and functional transformations to PC-12 cells. From their initial round or slightly elongated form, growing in suspension or as an adherent monolayer, NGF treatment propels them to exhibit extended neurites, signifying a profound shift towards a neuronal phenotype. These neurite outgrowths are crucial for cellular communication and interaction, reflecting the connectivity inherent in neuronal tissues.

Cancer Research and Cellular Mechanisms

In cancer research, the neuronal model offered by PC12 cells finds relevance owing to their tumorigenic origin. These cells are instrumental in probing cancer cell proliferation, invasion, and migration mechanisms. A 2020 study delved into the impact of Ferula assa-foetida extract on PC12 cell proliferation, shedding light on its potential therapeutic applications [7].

Stroke Research and Neuroprotective Strategies

Moreover, PC12 ADH cells, a variant adhering to culture surfaces, enable in-depth analyses of cell morphology and responses to stimuli like oxygen-glucose deprivation, serving as an essential model for stroke research and neuroprotective agent evaluation. The rapid doubling time of PC12 cells, ranging from 24 to 48 hours, underscores their efficacy in dynamic studies where cell proliferation rates are crucial.

Concluding Remarks on PC12 Cell Applications

In essence, PC12 cells and their derivatives embody a versatile model for a wide array of research applications spanning neurobiology, drug discovery, and cancer studies. Each application offers critical insights into cellular responses, paving the way for therapeutic innovations and advancing our understanding of complex biological processes.

Get the Robust & Versatile PC-12 Cell Line: Ideal for Neuronal Research & Easy to Culture

Highlighted Research Utilizing PC-12 Cells

PC-12 cells, derived from rat adrenal pheochromocytoma, are a widely used model in neurobiology, offering insights into neural differentiation, neurotoxicity, and more. Here are some notable studies featuring these cells:

Comprehensive Resources for PC-12 Cell Research

Access a variety of resources for in-depth understanding and manipulation of PC-12 cells, encompassing cell culture techniques, differentiation protocols, and more:

  • PC-12 Cell Differentiation Guide: This article provides a thorough overview of the PC12 cell NGF differentiation protocol, including methodologies for differentiation-related assays.
  • Video Tutorial on PC-12 Cell Transfection: Discover the intricacies of PC12 cell transfection through this detailed video guide, ideal for both novice and experienced researchers.
  • Mastering PC-12 Cell Culture: Delve into the nuances of subculturing PC-12 cells with this comprehensive protocol, essential for maintaining healthy cell cultures.

PC-12 Cell Line FAQs: Insights on Cultivation, Differentiation, and Research Applications

PC12 cells are derived from rat pheochromocytoma and are a type of cell line that exhibits neuronal characteristics when treated with specific growth factors such as NGF (nerve growth factor). They are commonly used as a model for neuronal differentiation and neurobiological studies.

PC12 cells are derived from rat pheochromocytoma and are a type of cell line that exhibits neuronal characteristics when treated with specific growth factors such as NGF (nerve growth factor). They are commonly used as a model for neuronal differentiation and neurobiological studies.

PC12 ADH cells are a variant of the standard PC12 cell line that are adherent, meaning they attach to the surface of the culture dish. This property makes them particularly useful for experiments requiring observation of cell morphology and cellular responses under various conditions.

The doubling time of PC12 cells can vary depending on the culture conditions, but it typically ranges from 24 to 48 hours. This rate can be influenced by factors such as the presence of growth factors like NGF or EGF, serum concentration, and the substrate on which the cells are grown.

NGF, or nerve growth factor, induces differentiation in PC12 cells, leading them to extend neurites, which are similar to the axons and dendrites of neurons. This makes PC12 cells a valuable model for studying neuronal growth, neurogenesis, and the molecular pathways involved in neural development.

PC12 cells subjected to oxygen-glucose deprivation can mimic the cellular stress conditions experienced during a stroke. This makes them a good model for studying the pathophysiology of stroke, examining neuroprotective compounds, and understanding cellular survival mechanisms under stress.

Yes, PC12 cells can be utilized in biosensing applications due to their ability to release neurotransmitters and express neuronal biomarkers. They can be used to develop biosensors that detect neurotoxicity or the efficacy of neuroprotective agents.

Epidermal growth factor (EGF) stimulates the proliferation of PC12 cells without inducing differentiation. It binds to EGF receptors on the cell surface, triggering pathways that lead to cell growth, making it useful for expanding the cell population in culture.

Yes, PC12 cells are suitable for studying neurodegenerative diseases as they can express markers and pathways relevant to neurodegeneration. They are often used in research on Alzheimer's disease, Parkinson's disease, and other neurodegenerative conditions.

Neuronal biomarkers in PC12 cells are significant because they can be used to monitor the differentiation and health of the cells, as well as to study the effects of neurotoxic agents and neuroprotective compounds in neurobiological research.

PC12 cells provide a platform to study and screen neuroprotective agents. By observing how these cells respond to different agents, particularly in conditions that simulate neuronal injury or stress, researchers can understand the effectiveness and mechanism of action of potential neuroprotective compounds.

PC-12 cells, derived from a transplantable rat pheochromocytoma of the adrenal gland, exhibit characteristics intrinsic to neurons, such as neurite outgrowth in response to nerve growth factor (NGF), making them an exemplary neuronal cell model. They are also used as cellular models to study pathophysiology and pharmacology due to their response to neurotrophic factors and susceptibility to neurotoxicity.

PC-12 cells are vital in cellular neurobiology for elucidating molecular mechanisms such as NGF receptors' interaction, dopamine metabolism, and cellular consequences of different stimuli. They aid in the investigation of the unfolded protein response, exocytosis, and vesicular trafficking—processes fundamental to neuronal cell death and survival.

Yes, PC-12 cells, especially adherent cells variants, are adaptable to a variety of culture conditions, including 3D cell culture and oxygen glucose deprivation. This adaptability makes them valuable for biosensing and biosensors development and in methodologies of cell biology for simulating different pathological states.

PC-12 cells originate from a tumor, making them relevant for studying cancer cell mechanisms, including proliferation influenced by factors like collagen IV and cell apoptosis. They provide insights into the cellular models of cancer pathophysiology, facilitating analyses in biomedical laboratories.

They are crucial in testing neuroprotective drugs and examining the cellular response to injury, with fgfs (fibroblast growth factors) in injury repair and cell apoptosis analysis being areas of significant interest. PC-12 cells are integral to drug discovery processes, particularly for their role in identifying compounds that can influence neuronal survival and neuroregeneration.

References

  1. Wu, C., et al., Icariin promotes the repair of PC12 cells by inhibiting endoplasmic reticulum stress. BMC Complement Med Ther, 2021. 21(1): p. 69.
  2. Sultan, N., et al., Dental pulp stem cells stimulate neuronal differentiation of PC12 cells. Neural Regeneration Research, 2021. 16(9): p. 1821.
  3. Ando, E., et al., Osteocalcin promotes proliferation, differentiation, and survival of PC12 cells. Biochemical and Biophysical Research Communications, 2021. 557: p. 174-179.
  4. Shanmuganathan, B., et al., Anti-amyloidogenic and anti-apoptotic effect of α-bisabolol against Aβ induced neurotoxicity in PC12 cells. European journal of medicinal chemistry, 2018. 143: p. 1196-1207.
  5. Fujita, K., P. Lazarovici, and G. Guroff, Regulation of the differentiation of PC12 pheochromocytoma cells. Environ Health Perspect, 1989. 80: p. 127-42.
  6. Abroudi, M., et al., Antiproliferative effects of Ferula assa-foetida's extract on PC12 and MCF7 cancer cells. Int J Biomed Engg Clin Sci, 2020. 6(3): p. 60-67.

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