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RBL-2H3 Cells - Significance of RBL-2H3 in Allergic Response and Immunological Studies

The RBL-2H3 cell line is a rat-derived basophilic leukemia cell line used in immunology research. It serves as a model system for studying mast cell physiology, behavior, and functions. These cells are also employed to study allergic reactions, immunological processes, and drug testing and development.

The article will provide insights into RBL-2H3 origin, general attributes, culture requirements, and research applications.

RBL-2H3 cells: Origin and general characteristics

Before using a basophil cell line in your research, you should know its origin and general attributes. This section of the article will inform you about RBL-2H3 cell basics. Such as, What are the RBL-2H3 mast cells? Why use RBL-2H3 cells? What are RBL-2H3 cells in rat basophilic leukemia? What is the RBL-2H3 morphology? Are RBL-2H3 cells immortal?

  • RBL-2H3 are basophilic leukaemia cells derived from Wistar rat basophil cells in 1978 at the National Institute of Dental Research Laboratory of Immunology.
  • RBL-2H3 cells possess c-kit receptor tyrosine kinase and mast cell protease II (RMCP-II) receptors, making them a potential mast model. Therefore, they are commonly called mast cells despite their origin from rat basophils [1].
  • They release histamine and other mediators upon activation and express high-affinity IgE receptors.
  • RBL-2H3 cells exhibit fibroblast-like morphology.

Microscopic visualisation of a mast cell.

RBL-2H3 Cells: Culturing information

This section will help you learn some key aspects for culturing the RBL-2H3 cell line. You will know: What is RBL-2H3 cell doubling time? What is the RBL-2H3 cell seeding density? What is the RBL-2H3 cell culture protocol? What is RBL-2H3 cell line freezing media?

Key Points for Culturing RBL-2H3 Cells

Population Doubling Time:

The RBL-2H3 mast cell doubling time is approximately 50- 60 hours.

Adherent or in Suspension:

The RBL-2H3 is an adherent cell line.

Sub-cultivation ratio:

For RBL-2H3 cells, the split ratio is kept between 1:2 to 1:4. Adherent cells are washed with magnesium- and calcium-free 1 x PBS solution. Passaging solution Accutase is added, and cells are kept at ambient temperature for 10 minutes to detach from the bottom of the culture vessel. Fresh media is added, and cells are centrifuged. Harvested cells are carefully resuspended in fresh medium and poured into new flasks containing growth medium.

Growth Medium:

EMEM media containing 10% FBS, 2 mM L-Glutamine, 2.2 g/L NaHCO3, and EBSS salt is used to culture the RBL-2H3 cells. Media should be replaced 2 to 3 times per week.

Growth Conditions:

RBL-2H3 cells are cultured in a humidified incubator set at 37 °C temperature and connected to a 5% CO2 source.


The cells are stored in the vapour phase of liquid nitrogen or at below -150 °C temperature in an electric freezer to protect the cell's viability for the long term.

Freezing Process and Medium:

CM-1 or CM-ACF freezing media is used to freeze RBL-2H3 cells using a slow freezing process. Briefly, this method permits a 1 °C decrease in temperature per minute and protects cells from shock.

Thawing Process:

RBL-2H3 cells are thawed in a pre-set water bath (37 °C) for approximately 60 seconds. Afterwards, cells are added to fresh culture medium and centrifuged. This step is essential to remove freezing media components. Next, the cell pellet is resuspended in a growth medium, and cells are dispensed into a flask for culturing.

Biosafety Level:

The RBL-2H3 cells should be maintained in biosafety 1 laboratories.

Adherent monolayers of RBL-2H3 cells at different confluence 20- and 10-fold magnification.

Advantages & Limitations of the RBL-2H3 Basophil Cell Line

RBL-2H3 cells are commonly utilized in immunological research. This section outlines their primary advantages and limitations.


  • Easy to Culture: RBL-2H3 cells can be easily cultured and maintained in the laboratory environment. This facilitates cost-effective and reproducible experimentation, making them a popular choice for initial studies in immunology.


  • Non-Human Origin: Originating from rat basophils, RBL-2H3 cells may not accurately mimic human biological processes, which can limit their applicability in human-specific research studies. This necessitates cautious interpretation of data when extrapolating results to human systems.
  • Simplified Mast Cell Model: While these cells provide a basic model for studying mast cell functions, they do not fully represent the complex nature of mast cell interactions within the human immune system. Consequently, they may not adequately model the multifaceted roles of mast cells in immune responses or disease conditions in vivo.

RBL-2H3 Cell Line: A Cornerstone in Immunological Research

Basophil and Mast Cell Research Utilizing the RBL-2H3 Cell Line

The RBL-2H3 cell line, derived from Rattus norvegicus, serves as a key model for studying the biology of basophils and mast cells. These rat mast cells provide essential insights into mast cell mediator release, critical to understanding allergic conditions such as allergic rhinitis. Through these cells, researchers explore cell receptor dynamics and the establishment of immunological synapses, which are central to the immune system's response to allergens.An interesting study published in 2019 used the RBL-2H3 cell line and investigated mechanisms behind pseudo-allergic reactions produced by Qingkailing injection. The study found that the PI3K-RAC1 signalling cascade partially elicits this allergic response in cells [2].

Immunological Synapse Dynamics in Allergy Research

Employed extensively in immunology research, RBL-2H3 cells are particularly effective for investigating immunological synapse dynamics. This aids in elucidating the communication processes of the immune system, with applications in studying both peripheral blood and peritoneal mast cells. Such research is vital for a comprehensive understanding of the immune response in both systemic and localized contexts.

Drug Screening and Toxicity Testing

In drug screening and testing, the responsiveness of RBL cells to various stimuli is exploited, including the study of how H2O2 inhibits IgE-mediated responses. These cells play an essential role in the development of treatments for diseases like streptococcal infections, where the mitis group streptococci inhibits mast cell activation. Besides this, researchers evaluate the toxic effects of various substances, including chemicals, drugs, and nanoparticles, using this RBL 2H3 mc model. Such as a recent study (2022) assessed the cytotoxicity of polystyrene microplastics on RBL-2H3 cells. They found that microplastics damage RBL-2H3 cell organelles and promote cell death [3]. Another 2021 study evaluated the anti-allergic and anti-inflammatory potential of a natural product, neferine, using the RBL 2H3 mc model. The study revealed the compound's good anti-allergic and anti-inflammatory properties [4].

Advanced Methodologies in Immunological Measurement

The consistent and measurable mediator release from RBL-2H3 cells makes them ideal for convenient fluorimetry, facilitating precise and accurate measurements crucial for disease study and the assessment of therapeutic agents.

The RBL-2H3 cell line, with its origins in Rattus norvegicus, is an invaluable asset in both fundamental and applied research within the field of immunology. It offers profound opportunities for advancing our knowledge and treatment of immunological diseases.

RBL-2H3 Cell Line for Advanced Immunological Research

RBL-2H3 cells: Research publications

The following are some exciting research publications featuring the RBL-2H3 mast cells:

Inhibitory activity of narirutin on RBL-2H3 cells degranulation

This research paper was published in the Immunopharmacology and Immunotoxicology (2021). The study proposed that narirutin, a natural compound, exerts inhibitory effects on RBL 2H3 degranulation via regulating NF-κB, MAPK, and the tyrosine kinase signalling pathway.

Effects of apigenin on RBL-2H3, RAW264. 7, and HaCaT cells: anti-allergic, anti-inflammatory, and skin-protective activities

This research in the International Journal of Molecular Sciences (2020) proposed that the apigenin compound significantly suppresses allergic and inflammatory responses of RBL-2H3 and RAW264.7 cells. Therefore, it may serve as a potential agent for combating immune-related diseases.

Anti-inflammatory and anti-allergic effects of saponarin and its impact on signaling pathways of RAW 264.7, RBL-2H3, and HaCaT Cells

This research paper in the International Journal of Molecular Sciences (2021) evaluated the anti-allergic and anti-inflammatory effects of a natural product, saponarin, using different cell lines, including RBL-2H3.

Inhibitory Effect on β -Hexosaminidase Release from RBL-2H3 Cells of Extracts and Some Pure Constituents of Benchalokawichian, a Thai Herbal Remedy, Used for Allergic Disorders

This Evidence-Based Complementary and Alternative Medicine (2014) study found that Benchalokawichian extracts and some active constituents exert potential inhibitory effects on RBL 2H3 ß-hexosaminidase release.

Spirulina maxima peptides suppress mast cell degranulation via inactivating Akt and MAPKs phosphorylation in RBL-2H3 cells

This paper was published in 2018 in the International Journal of Biological Macromolecules. The study stated that spirulina maxima, a natural product, inhibits RBL 2H3 degranulation by preventing MAPKs and AKT phosphorylation.

Resources for RBL-2H3 cell line: Protocols, Videos, and More

RBL-2H3 is a widely used mast cell line. The available resources covering RBL-2H3 culturing and transfection protocols are mentioned here:

  • RBL 2H3 MC model: This research article comprises protocols for maintaining RBL-2H3 cultures and for RBL-2H3 transfection.

Here are some resources explaining the RBL-2H3 cell culture protocol:

  • RBL-2H3 cells: This website is beneficial for you to learn cell culture protocols for the RBL-2H3 cell line. Besides, it comprises information about RBL 2H3 cell media and culturing conditions.

RBL-2H3 Cell Line: Essential FAQs for Researchers

RBL-2H3 cells are a cell line originally derived from the rat basophilic leukemia and are commonly used as a model to study mast cell biology, particularly in the context of allergic reactions and immunological research. These cells are valuable for analyzing basophil and mast cell mediator release, cell receptor dynamics, and the formation of immunological synapses.

Yes, upon activation, RBL-2H3 cells are known to release histamine and other inflammatory mediators. This characteristic makes them highly useful for studies related to allergic responses, as histamine release is a key feature of allergic reactions and inflammatory processes.

The doubling time of RBL-2H3 cells varies depending on culture conditions but is generally around 24 to 36 hours. However, optimal growth conditions such as temperature, pH, and nutrient availability can influence the doubling time.

While RBL-2H3 cells are of rat origin and there are inherent differences between species, they are still widely used as a model to study mechanisms relevant to human allergic responses. However, results from RBL-2H3 cell studies should be carefully interpreted when extrapolating to human systems.

H2O2, or hydrogen peroxide, has been shown to inhibit IgE-mediated degranulation in RBL-2H3 cells. This effect is significant for researchers studying the modulation of allergic reactions at a cellular level.

Yes, RBL-2H3 cells are extensively used for drug screening, especially for compounds that target allergic and inflammatory pathways. They are used to evaluate the efficacy and safety of potential therapeutic agents, as well as their anti-allergic and anti-inflammatory properties.


  1. Passante, E. and N. Frankish, The RBL-2H3 cell line: its provenance and suitability as a model for the mast cell. Inflamm Res, 2009. 58(11): p. 737-45.
  2. Li, Q., et al., Pseudo-allergic reaction caused by Qingkailing injection partially via the PI3K-Rac1 signaling pathway in RBL-2H3 cells. Toxicology Research, 2019. 8(3): p. 353-360.
  3. Liu, L., et al., Polystyrene micro (nano) plastics damage the organelles of RBL-2H3 cells and promote MOAP-1 to induce apoptosis. Journal of Hazardous Materials, 2022. 438: p. 129550.
  4. Chiu, K.-M., et al., Anti-allergic and anti-inflammatory effects of neferine on RBL-2H3 cells. International Journal of Molecular Sciences, 2021. 22(20): p. 10994.

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