P19 Cells - Embryonal Carcinoma Research using

P19 Cells

P19 is a murine embryonal carcinoma cell line. It is widely used in biomedical research, mainly for studying developmental biology, stem cell biology, cell differentiation, and drug screening. As P19 cells have differentiation ability, they can be useful in investigating complex biological processes such as tissue formation and early embryonic development. In this article, we will discuss the fundamentals of mouse-derived P19 cells. Particularly, we will talk about:

General characteristics and origin of P19 cells
Culturing information on P19 cell line
P19 cell line: Advantages & disadvantages
Research applications of P19 cells
P19 cells: Research publications
Resources for P19 cell line: Protocols, Videos, and More

1. General characteristics and origin of P19 cells

Knowing about a cell line's general characteristics and origin is essential before you start working with it. This section will cover the following: What is P19 cell line? What is the size of the P19 cell? What is the origin of P19 cells?

P19 is a type of pluripotent embryonal carcinoma cells originally obtained from teratocarcinoma developed in a C3H/He mouse. The cell line was first established in 1982 by McBurney and Rogers.
P19 cells can continuously grow in a serum-supplemented culture media. They can be differentiated into other cell types when exposed to non-toxic drugs such as retinoic acid and dimethyl sulfoxide (DMSO) [1].
These mouse carcinoma cells possess an epithelial-like morphology.
The P19 cell line has a euploid male karyotype (n=40; XY).

Modeling mitosis of embryonic stem cells, magnified in the microscope.

2. Culturing Information on P19 Cells

The P19 cell line is widely cultured in research laboratories due to its unique characteristics. Its culturing is easy and manageable. This section has mentioned all the key information you need to maintain and grow P19 cell culture. We will know: What is the doubling time of P19 cells? How do you culture P19 cell line? Is P19 an adherent cell line?

Key Points for Culturing P19 Cells

Doubling Time:	The doubling time reported for the P19 cell line is approximately 2 to 3 days.
Adherent or in Suspension:	P19 embryonic carcinoma cell line is adherent.
Subculturing ratio:	P19 cells should be subcultured every 48 hours, and a split ratio of 1:10 should be maintained for these cells. Adherent cells are washed with 1 X phosphate buffer saline and incubated with Accutase until the cells dissociate. Cells are added with culture media and harvested through centrifugation. The collected cells are carefully resuspended and dispensed into new flasks.
Growth Medium:	DMEM/Ham's F12 media containing 5% Fetal bovine serum, 3.1 g/L Glucose, 1.6 mM L-Glutamine, 1.0 mM Sodium pyruvate, 15 mM HEPES and 1.2 g/L NaHCO3 is used to culture P19 cells.
Growth Conditions:	A humidified incubator set at 37°C with a 5% CO2 supply is essential to growing and culturing the P19 embryonic carcinoma cell line.
Storage:	Frozen P19 cell vials should be stored at below -150°C temperature in a freezer or the vapour phase of liquid nitrogen to maintain the viability of cells for the longer term.
Freezing Process and Medium:	CM-1 or CM-ACF media can be used to freeze P19 cells using a slow freezing method that protects cells from any shock and preserve their viability.
Thawing Process:	Frozen P19 cells can be thawed in a 37°C water bath by rapidly agitating a vial for 40 to 60 seconds. Cells are added with fresh media and centrifuged to remove freezing media elements. The cell pallet is again resuspended, and cells are poured into the new flask for growth.
Biosafety Level:	Biosafety level 1 laboratory settings are required for the P19 cell line.

Adherent and semi-confluent layer of P19 cells at 10× and 20× magnification.

3. P19 cell line: Advantages & Disadvantages

This section will discuss the advantages and disadvantages of the P19 cell line.

Advantages

The main advantages of the P19 cell line are:

Differentiation potential

P19 cells can differentiate into various cell types, including cardiomyocytes, neurons, microglial cells, etc. They require nontoxic drugs to differentiate, such as retinoic acid and dimethyl sulfoxide (DMSO). Retinoic acid induces the development of neurons, microglia, and astroglia, whereas DMSO initiates beating cardiomyocytes and smooth muscle cell development from P19 cells. Thus, P19 cells can be useful in studying cell differentiation and developmental processes.

Model system

Pluripotent embryonic carcinoma cell line P19 is a useful model for studying early embryonic development. Researchers use P19 cells to elucidate cell signaling pathways and cellular and molecular mechanisms involved in these processes.

Disadvantages

The disadvantage associated with the P19 cell line is:

Murine origin

P19 is a murine embryonal carcinoma cell line. Therefore, the study findings from these cells may not fully relate to human biology and processes.

4. Research applications of P19 cells

P19 cells have several research applications due to their differentiation ability and relevance to developmental biology and stem cell research. Some of the important research applications of P19 embryonic carcinoma cells include:

Cell differentiation studies: As we know, P19 cells can differentiate into neurons, microglial, smooth muscle cells, and cardiomyocytes; thus are widely used to study cell differentiation processes. In addition, it helps research investigate neural and cardiac development and underlying mechanisms. A study conducted in 2018 found that reactive oxygen species (ROS) direct the differentiation of P19 cells into specific cell types and prevent the induction of others [3]. Another study explored the retinoic acid-mediated neural differentiation process and found the involvement of the PI3K/Akt/GSK3β signalling pathway [4].
Development biology: P19 cells are an invaluable model for studying early embryonic development. They help researchers understand complex biological processes, such as tissue formation during embryo development. The research used P19 cells and studied contributing molecular factors for ventricular septal defect (VSD) formation. The findings revealed that a long noncoding RNA SNHG6 contributes to VSD by negatively regulating miRNA-101 and activating the Wnt/β-catenin pathway [5].
Drug testing: P19 mouse embryonic carcinoma cell line is also used to screen potential drug candidates. A study used differentiated P19 cell neurons and investigated the neuroprotective acetylcholinesterase Inhibitory effects of synthetic L-Dopa and Mucuna pruriens seed aqueous extract. The results showed that plant extract exhibited promising results compared to L-Dopa [6].

5. P19 cells: Research publications

This article section will cover a few interesting research publications featuring P19 cells.

Novel evidence that pituitary sex hormones regulate migration, adhesion, and proliferation of embryonic stem cells and teratocarcinoma cells

This article was published in Oncology Reports in 2017. The study proposed that pituitary sex hormones drive teratocarcinoma cell lines' adhesion, proliferation, and migration, including P19 cells.

The long non-coding RNA uc. 4 influences cell differentiation through the TGF-beta signaling pathway

This publication in the Experimental & Molecular Medicine journal (2018) used P19 cells and studied the function of long noncoding RNA uc.4. The findings revealed that uc.4 affects cell differentiation via modulating the TGF beta signalling pathway.

The combined effects of three‐dimensional cell culture and natural tissue extract on neural differentiation of P19 embryonal carcinoma stem cells

This research article was published in 2018 in the Journal of Tissue Engineering and Regenerative Medicine. The study found that natural brain tissue extract and 3D cell culture can expedite P19 embryonic carcinoma cells' differentiation into neural cells.

In vitro differentiation induction of embryonal carcinoma stem cells into insulin-producing cells by Cichorium intybus L. leaf extract

This study was published in the Journal of Ethnopharmacology in 2020. The study proposed that Cichorium intybus L. leaf extract can induce differentiation in P19 embryonic carcinoma cells into insulin-producing pancreatic β cells.

Mucuna pruriens Seed Aqueous Extract Improved Neuroprotective and Acetylcholinesterase Inhibitory Effects Compared with Synthetic L-Dopa

This research was published in Molecules (2022). This study explored the neuroprotective and acetylcholinesterase inhibitory effects of Mucuna pruriens seed extract on P19 cell neurons.

6. Resources for P19 cell line: Protocols, Videos, and More

The following are a few resources on P19 cells.

P19 cells neuronal differentiation protocol: This article contains P19 cells neural differentiation protocol and other useful information about P19 cell differentiation.
P19 cell transfection: This link will help you learn the P19 cell transfection protocol.

The following link contains the p19 cell culture protocol.

P19 cells: This website contains all useful information about the P19 cell line, including its culture condition, P19 cell media, cell splitting, and much more.

References

McBurney, M.W., P19 embryonal carcinoma cells. Int J Dev Biol, 1993. 37(1): p. 135-40.
Bressler, J., et al., P19 Embryonic Carcinoma Cell Line: A Model To Study Gene–Environment Interactions. Cell Culture Techniques, 2011: p. 223-240.
Pashkovskaia, N., U. Gey, and G. Rödel, Mitochondrial ROS direct the differentiation of murine pluripotent P19 cells. Stem Cell Research, 2018. 30: p. 180-191.
Fu, F., et al., All‐trans‐retinoid acid induces the differentiation of P19 cells into neurons involved in the PI3K/Akt/GSK3β signaling pathway. Journal of Cellular Biochemistry, 2020. 121(11): p. 4386-4396.
Jiang, Y., et al., Long noncoding RNA SNHG6 contributes to ventricular septal defect formation via negative regulation of miR-101 and activation of Wnt/β-catenin pathway. Die Pharmazie-An International Journal of Pharmaceutical Sciences, 2019. 74(1): p. 23-28.
Kamkaen, N., et al., Mucuna pruriens seed aqueous extract improved neuroprotective and acetylcholinesterase inhibitory effects compared with synthetic L-dopa. Molecules, 2022. 27(10): p. 3131.