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.

This article contains useful information about the PC-12 cell line to ease your work. It will mainly focus on the following:

General information and origin of the PC-12 cell line
Culturing of PC-12 cells
PC-12 cell line: Advantages & Limitations
Research applications of PC-12 cell line
Publications Featuring PC-12 Cells
Resources for PC-12 Cell line: Protocols, Videos, and More

1. 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).

PC12 and PC12 ADH

PC12 ADH is a subline of the PC12 parental cell line. The main difference between these two cells is that PC12 cells are irregularly shaped floating cell clusters that grow in suspension and poorly adhere to non-coated surfaces. In contrast, PC12 ADH is adapted and has good adherence ability to plastic surfaces [2].

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

2. 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 10⁴cells/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.

3. 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 cover some key advantages and limitations associated with the cell line.

Advantages

The advantages of the PC 12 cell line are:

Easy to Culture	Robust and relatively easy to culture
Neuronal Differentiation	They can be induced to differentiate into neuron-like cells and are an excellent model for studying neural development, differentiation, and processes associated with neurogenesis.

Limitations

The limitations of the PC-12 cell line are:

Non-Human Origin

They are derived from rat pheochromocytoma, limiting their applicability for certain human-specific research studies. It may not fully represent human neural physiology.

4. Research applications of PC-12 cell line

The PC 12 cell line has a wide range of research applications. A few promising applications are listed in this section:

Neurobiology studies: The PC 12 cell line offers a range of applications in neurobiology studies. It is an invaluable cell model for exploring neural differentiation, neurotrophic factor responses, and neurotoxicity. An interesting study investigated the potential effects of dental pulp stem cells and derived factors on the differentiation, survival and growth of PC12 cells [3]. A similar study found that Osteocalcin, a calcium-binding protein, encourages survival, growth and differentiation of neural cell line (PC-12) [4].
Drug Screening: PC12 cells are valuable research tools for screening potential drugs and compounds for their neuroprotective or neurodegenerative potential. This may aid in discovering novel and effective drugs against neurodegenerative disorders. Such a study explored the neuroprotective effects of a natural product called alpha-bisabolol, using a PC12 cells Parkinson's disease model. The results showed that α-bisabolol exerts anti-amyloidogenic, anti-apoptotic effects and modulates amyloid beta-induced neurotoxicity in PC12 cells [5].
Cancer studies: PC12 cells neuronal model also offers applications in cancer research due to their tumour origin [6]. Therefore, they are used to study cancer cell proliferation, invasion, and migration. PC12 cancer applications were explored in 2020. The study explores the potential effects of Ferula assa-foetida extract on PC-12 cell proliferation [7].

5. Publications Featuring PC-12 Cells

The following are some research publications featuring the PC12 neuronal cells:

Caffeic acid phenethyl ester (CAPE) protects PC12 cells from cisplatin-induced neurotoxicity by activating the NGF-signaling pathway

This study published in Neurotoxicity Research (2017) explored the neuroprotective effect of caffeic acid phenethyl ester (CAPE) in the cisplatin-induced peripheral neuropathy cell model (PC-12).

Krill oil protects PC12 cells against methamphetamine-induced neurotoxicity by inhibiting apoptotic response and oxidative stress

This study was published in Nutrition Research (2018). It proposed that krill oil has the potential to impede methamphetamine-induced neurotoxicity in the PC-12 cell line by repressing oxidative damages and apoptotic responses.

Apios americana Medik flowers extract protects PC12 cells against H2O2-induced neurotoxicity via regulating autophagy

This research article in Food and Chemical Toxicology (2019) proposed that the Apios americana Medik flowers extract prevents hydrogen peroxide (H2O2) induced damage in the PC 12 cell line via regulating autophagy.

Angiotensin II triggers autophagy and apoptosis in PC12 cell line: an in vitro Alzheimer's disease model

This research was published in Brain Research (2019). The study explored that angiotensin II, a medication, induces apoptosis and autophagy in PC12 neuronal cells (Alzheimer's disease cell model).

Evaluation of toxicity of glycerol monooleate nanoparticles on PC12 cell line

This research article published in the International Journal of Pharmaceutics (2018) evaluated the glycerol monooleate nanoparticles toxicity using PC12 cell line.

6. Resources for PC-12 Cell line: Protocols, Videos, and More

Many resources are available on the PC-12 neuronal cell line. This section will mention a few resources covering these neural cells' cell culture and transfection protocol.

PC-12 cell differentiation: This article will explain the PC12 cells NGF differentiation protocol. Moreover, the publication comprises a protocol for differentiation-related assays, including pc12 cells neurite outgrowth assay.
Transfection of PC12 cells: This video tutorial is a step-wise guide to learning transfection protocol for PC12 cells.

The PC12 cell culture protocol is mentioned here:

PC 12 cell line: This document will help you learn the protocol for subculturing the PC 12 cell line.

References

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.
Wiatrak, B., et al., PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions. Cells, 2020. 9(4).
Sultan, N., et al., Dental pulp stem cells stimulate neuronal differentiation of PC12 cells. Neural Regeneration Research, 2021. 16(9): p. 1821.
Ando, E., et al., Osteocalcin promotes proliferation, differentiation, and survival of PC12 cells. Biochemical and Biophysical Research Communications, 2021. 557: p. 174-179.
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.
Fujita, K., P. Lazarovici, and G. Guroff, Regulation of the differentiation of PC12 pheochromocytoma cells. Environ Health Perspect, 1989. 80: p. 127-42.
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.