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. By reading this article, you will thoroughly understand:

  1. RBL-2H3 cells: Origin and general characteristics
  2. RBL-2H3 cell line: Culturing information
  3. Advantages & Limitations of RBL-2H3 cells
  4. Applications of RBL-2H3 cell line
  5. RBL-2H3 cells: Research publications
  6. Resources for RBL-2H3 cell line: Protocols, Videos, and More

1.      RBL-2H3 cells: Origin and general characteristics

Before using a 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.

2.      RBL-2H3 cell line: Culturing information

Culturing information about a cell line is crucial for its proper handling and maintenance in research laboratories. This section will help you learn key points 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.

3.      Advantages & Limitations of RBL-2H3 cells

RBL-2H3 cells are generally employed in immunology studies. Like other cells, they also offer some advantages and have limitations. A few of these are listed in this section.


The main advantages of the RBL-2H3 cell line include:

Easy Culturing

These cells are easy to culture and maintain in the laboratory, allowing for cost-effective and reproducible experimentation.



Here are some limitations associated with the RBL-2H3 cell line:

Non-Human Origin

RBL-2H3 cells were developed from rat basophils, limiting their relevance for certain human-specific research studies.

Simplified Mast Cell Model

RBL-2H3 cells present a simplified model of mast cells, potentially not fully recapitulating the complexity of mast cells and their interactions with other immune system cells in vivo.


4.      Applications of RBL-2H3 cell line

The RBL-2H3 cell line has numerous applications in research. Here, we have talked about some significant research uses of this basophilic leukaemia cell line.

  • Immunology Research: RBL-2H3 cells release histamine and other inflammatory mediators upon activation and thus are ideally used to study allergic reactions and inflammatory responses. Researchers also employ these cells to investigate molecular mechanisms underlying allergic responses and immune reactions. 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].
  • Drug Screening and testing: The RBL-2H3 cells are commonly used in screening drugs and compounds for their therapeutic effects on allergic and inflammatory responses. This is imperative for identifying and developing potential treatments for different allergic disorders. 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].

5.      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.

6.      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.


  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.