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.
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).
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
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Doubling Time: |
The doubling time reported for the P19 cell line is approximately 2 to 3 days. |
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Adherent or in Suspension: |
P19 embryonic carcinoma cell line is adherent. |
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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. |
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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. |
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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. |
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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. |
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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. |
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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. |
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Biosafety Level: |
Biosafety level 1 laboratory settings are required for the P19 cell line. |
P19 cell line: Advantages & Disadvantages
This section will discuss the advantages and disadvantages of the P19 cell line.
Advantages
- Differentiation potential: P19 cells can differentiate into various cell types, including cardiomyocytes, neurons, and microglial cells. They require nontoxic drugs for differentiation, such as retinoic acid and dimethyl sulfoxide (DMSO). Retinoic acid induces the development of neurons, microglia, and astroglia, whereas DMSO initiates the development of beating cardiomyocytes and smooth muscle cells. Thus, P19 cells are useful in studying cell differentiation and developmental processes.
- Model system: The pluripotent embryonic carcinoma cell line P19 is a valuable model for studying early embryonic development. Researchers utilize P19 cells to elucidate cell signaling pathways and the cellular and molecular mechanisms involved in these processes.
Disadvantages
- Murine origin: P19 is a murine embryonal carcinoma cell line. Consequently, the findings from studies using these cells may not fully translate to human biology and processes.
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].
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P19 cells: Research publications
This article section will cover a few interesting research publications featuring P19 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.
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.
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.
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.
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.
Exploring the P19 Cell Line: Frequently Asked Questions
Cardiac differentiation in P19 cells refers to their ability to develop into cardiomyocyte-like cells under specific conditions, such as the presence of dimethyl sulfoxide (DMSO). This property makes them useful for studying cardiomyocyte differentiation and cardiac muscle cell development.
Undifferentiated P19 cells can be induced to differentiate into various cell types including neuronal and cardiac cells. This transition is influenced by the culture conditions and the application of differentiation agents like retinoic acid for neurons and DMSO for cardiac muscle cells.
Neuronal differentiation markers in P19 cells include genes associated with neurogenesis, such as those coding for neurotransmitters and specific neuronal marker genes. These markers help in identifying and characterizing the neuronal phenotype.
Ion channel expression in P19 cells can be studied by examining the mRNA expression of ion channel genes and through functional assays to assess ion channel activity. This is crucial for understanding how ion channels contribute to cell functions and neurogenesis.
P19 cells express clock genes, which regulate circadian rhythms affecting various cellular functions. Researching these genes helps understand the interplay between the circadian clock and cellular processes such as cell cycle regulation and metabolism.
Yes, under certain conditions, P19 cells can differentiate into glial cells, expanding their use in studies on neurogenesis and the role of glial cells in the central nervous system.
Characterization of the neurotransmitter phenotype in P19 cells involves identifying the specific neurotransmitters they produce, which is essential for studies on neurotransmission and neural function.
Aggregates of P19 cells are used to model more complex, three-dimensional structures that more closely mimic tissues, enhancing studies on cellular interactions and tissue development.
Multipotent P19cl6 cells are a subclone of P19 cells capable of differentiating into multiple cell types, including both neuronal and cardiac cells. Their multipotency makes them valuable for various research applications.
The effect of external factors on P19 cells can be studied using various assays, including colony assays for characterizing growth and differentiation, as well as electrophysiological assessments to understand changes in cell function.
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.