Jurkat E6.1 Cell Line

Jurkat E6.1 is a clone of the Jurkat cell line, an immortalized human T-lymphocytes cell line derived from peripheral blood. First established in 1977, they are used in research related to T-cell receptor signalling, T-cell and other lymphocyte adoptive therapies against cancer, blood-related proto-oncogenes research, cell survival and cell differentiation.

In this article, we will include all the information and pre-requisites you need to understand before starting your work on Jurkat E6.1 cloned cell lines which include:

Origin, History and General Information
Cell Culture Characteristics
Applications of Jurkat E6.1 Cell Lines
Research Publications
Advantages and Limitations
Resources: Protocols, Videos & More

1. Origin, History and General Information

Here is all the necessary information related to the history, origin, and general characteristics of the Jurkat E6.1 cell line and their handling in cell culture.

Jurkat E6.1 is an immortalized human T-lymphocyte cell line. This cloned cell line is a derivative of the Jurkat cell line obtained from the peripheral blood of a 14-year-old boy suffering from acute lymphoblastic leukaemia in 1976. After isolation from blood, these cell line was named JM initially, as JM and Jurkat indicate that these cell lines are isolated from the same patient. The Jurkat E6.1 cell line was cloned from cell derivatives known as Jurkat-FHCRC.
E6.1 cell lines are derived from leukaemia; these cell lines are used to increase the expression of IL-2 via simulation with monoclonal antibodies or lectin. They express T-cell antigen receptors; therefore, these cell lines can be used in the study of immune system disorders and immune-oncology research.
Cell Morphology of Jurkat E6.1 cells: Jurkat E6.1 cells were isolated from a leukaemia patient and have altered morphology as compared to T-Lymphocytes making them immortal cell lines. The lymphoblast cell forms when naïve lymphocytes are activated by antigen presentation by antigen-presenting cells. This results in increased growth in the nucleus, cytoplasm, mRNA and proteins.
Cell size: Jurkat E6.1 cell lines are cloned immortalized with more commonly the diameter ranging from 10-16 μm (the diameter may vary according to the type of suspension). These cell lines have a highly spherical cell shape.
Genome and Chromosome number: These cell lines are pseudodiploid cell lines of human T-lymphocytes, with their model chromosome chromosome 46. These also show that the modal chromosome is present in about 74% and shows a polyploidy rate of about 5.3%. The karyotype studies showed a karyotype with 46, X, Y, -2, -18, del (2) (p21p23), del (18) (p11.2). X and Y chromosomes typically have a regular copy in most cell lines.
Jurkat cell lines vs E6.1 Jurkat Cell lines: Jurkat cell lines are the primary parent cell lines first isolated from the acute lymphoblastic leukaemia patient. However, the E6.1 Jurkat cell lines are cloned derivate of Jurkat-FHCRC cell lines.

T cells fighting cancer: CAR T cell immunotherapy.

2. Cell Culture Characteristics

Key Points for Culturing B16 Cells

Population doubling time	The population doubling time of Jurkat cell lines is approximately 20.7 hours, allowing for relatively rapid cell development.
Adherent or non-adherent	E6.1 Jurkat cell lines grow in a non-adherent mode, existing in suspension either as single-layer growth or as free-floating clumps.
Seeding Density	The recommended seeding density for E6.1 Jurkat cell lines is 1.0-2.0 x 105 viable cells/mL to ensure healthy attachment and optimal cell growth.
Growth Medium	The required growth medium for E6.1 Jurkat cell lines is ATCC-formulated RPMI-1640 Medium (ATCC 30-2001). The complete medium is prepared by adding the specified components, including fetal bovine serum (ATCC 30-2002), to achieve a final concentration of 10%.
Growth Conditions	E6.1 Jurkat cell lines can be cultured in a suitable incubator with 5% air and CO2 availability using RPMI-1640 medium and fetal bovine serum.
Storage conditions	Long-term storage of Jurkat cell lines in a frozen culture state should be stored at -130 degrees Celsius or in liquid nitrogen in a vapour phase. Storing them at -70 degrees Celsius can lead to a loss of cell viability.
Freezing	An aliquot of approximately 0.5-1.0 mL of cell line suspension is stored in cryogenic tubes to maintain the cell lines. These tubes are then placed in a cryostorage container at -70 degrees Celsius. The tubes are transferred to a liquid nitrogen tank and stored in the vapor phase the following day.
Thawing	To thaw the Jurkat cell lines, the cell suspension vials should be placed in a water bath at 37 degrees Celsius. The suspension can be thawed for 2 minutes with gentle agitation. It is essential to keep the vial cap outside the water bath to prevent contamination of the cell lines.
Biosafety level	The growth of Jurkat cell lines requires a biosafety level of BSL-1 in the laboratory setting, ensuring basic safety precautions are followed.

Jurkat E6.1 cells growing in clusters at 10x and 20x magnification.

3. Applications of Jurkat E6.1 Cell Lines

Jurkat E6.1 cell line is most prominently used in research and studying disorders linked to the immune system and its associated cancers. Moreover, it has been suitable for the study of some complexities related to HIV infection since the CD4 T cell receptor is necessary for retroviral entry.

Some key areas where these cell lines are utilized are discussed below:

Model System for T Cell Signaling and Apoptotic Pathways: This T-cell leukaemia-derived cell line is used extensively in research involving TCR signalling cascade. Jurkat E6.1 releases more IL-2 and IL-8 that subsequently help in T-Cell activation and trigger the inflammatory pathways. This cell line is also known for its hyperphosphorylated Pyk2 and augmented production of Vav1 that crucially influences actin cytoskeletal rearrangement in T cells. TCR segregation in Jurkat Cell line E6-1 can also be improved by forceful expression of LFA-1 that consequently enhances TCR signalling and IL-2 expression. Cell proliferation and cell cycle studies are also conducted in Jurkat E6.1 cell lines. They have been preferred in studying apoptotic pathways and DNA damage induction by compounds derived from marine and plant sources. A study in 2013 evaluated impact of shark epigonal conditioned media on human cells by using these cell lines. Similarly, the repressive impact of plant-derived hirsutine on cell proliferation was studied in Jurkat E6.1 cell lines. Often anticancer studies also utilize Jurkat E6.1 to evaluate their selected targets. The influence of marine neurotoxins such as Brevetoxins (PbTx) has also been studied in these cell lines.

HIV Studies: One of the most significant applications of Jurkat E6.1 cell lines is their ability to serve as a suitable model for HIV studies due to their property of choosing different signalling events that induce TCA. Jurkat E6.1 cell lines have not only been used in understanding HIV activation but also in dissecting mechanisms behind HIV latency. HIV replication and infection have vastly been studied in these cell lines where they can be subjected to gene editing and gene knockout, as was done by a study in 2021. The impact of drug delivery on HIV infection, such as long-acting efavirenz (Efa)–enfuvirtide (Enf) Co-loaded polymer–lipid hybrid nanoparticles (PLN) on viral entry utilized Jurkat E6.1 cell lines. CRISPR/Cas9 engineered Jurkat E6.1 has been used multiple times for studying mechanisms involved in HIV entry, replication and infection.

4. Research Publications

Here are a few notable examples from the large collection of research, including Jurkat E6.1 cell lines.

The selenium metabolite methylselenol regulates the expression of ligands that trigger immune activation through the lymphocyte receptor NKG2D

Using Jurkat E6.1, incubated in different selenium compounds to study ligand expression and surface expression of MICA/B.

Caracasine acid, an ent-3,4-seco-kaurene, promotes apoptosis and cell differentiation through NFkB signal pathway inhibition in leukemia cells.

Understanding caracasine acid-induced cytotoxicity in human T cell lines such as Jurkat E6.1 and HL-60.

Evidence of In Vitro Development of Drug Resistance to Azidothymidine in T-Lymphocytic Leukemia Cell Lines (Jurkat E6–1/AZT-100) and in Pediatric Patients with HIV-1 Infection

Utilized Jurkat E6.1 incubated long-term with AZT drug to study Azidothymidine (AZT) resistance in HIV-1 infected patients.

Jurkat E6.1 Cell Line Studies Regarding The Effects of Some Bio-Indols on The Membrane Fluidity

This research utilized a polarized fluorescence analysis system to understand the impact of bio-indols, including serotonin, tryptophan and melatonin, on cell membrane fluidity in Jurkat E6.1 cell lines.

Molecular mechanism of apoptosis induction in Jurkat E6-1 cells by Tribulus terrestris alkaloids extract

Tribulus terristris-derived alkaloid extract induced apoptosis in Jurkat E6.1 cells to understand the underlying mechanism of action.

Increasing LFA-1 Expression Enhances Immune Synapse Architecture and T Cell Receptor Signaling in Jurkat E6.1 Cells

T Cell receptor signalling in Jurkat E6.1 cells was enhanced by forcefully increasing LFA-1 expression.

5. Advantages and Limitations

Like other cell lines, Jurkat E6.1 has its advantages and limitations.

Advantages

There are many reasons why it is preferred in these applications:

Mode of Growth
Jurkat E6.1 is a suspension and non-adherent cell line, meaning it grows suspended and does not adhere to the culture surface.
Expression of Genes
Jurkat E6.1 cell line expresses genes encoding interleukin-2 (IL‑2) and CD3. Upon stimulation with monoclonal antibodies, lectins, or phorbol esters against the T3 antigenic complex, these cells have been shown to produce amplified IL-2.
Expression of Antigens
Jurkat E6.1 cell line expresses CD3 antigen, associated with T cell activation and signalling.
Expression Markers
The cell line expresses the T cell antigen receptor (TCR) markers, which are crucial in T cell recognition and immune response.
Applicability for Transfection
Jurkat clone E6.1 cell line is suitable for host transfection experiments, allowing for the introduction of exogenous genetic material into the cells for various research purposes.

Limitations

After advantages, let us go through the limitations associated with the usage of Jurkat E6.1 cell lines:

Cell Transfection with Viral Vector
Transfecting Jurkat E6.1 cells is challenging due to their T-cell origin. Typically, expensive and time-consuming viral vectors are required for genetic modification. Non-viral vectors have shown low transfection efficiency and significant toxicity in Jurkat cells.
Bacterial Contamination
Jurkat E6.1 cell cultures should be monitored for potential bacterial contamination to ensure the integrity of experimental results.
Viral Contaminations
Jurkat E6.1 cell lines express various chemokine receptors and low levels of CD4, facilitating viral entry. This makes them susceptible to T cell-infecting viruses such as HTLV and HIV.

6. Resources: Protocols, Videos & More

This section will cover the methods and protocols for cell culturing and transfection, as well as educational videos related to this cell line:

Cell Culture Protocols

Here are some basic culture procedures to follow for this cell line:

Cell Culture SOP, Propagation of Jurkat: The University of California also provided a method on their USCS Genome browser for culturing Jurkat clone E6-1.
Jurkat, Clone E6-1 Culture and Handling: The Elabscience included culture handling protocol on their product description site.

Transfection Protocols

Here is the list of different transfection protocols available:

Jurkat Cells Transfection Protocol Lipofectamine LTX Reagent: A Jurkat Cell transfection protocol using Lipofectamine LTX Reagent, from ThermoFisher.
Amaxa™ 4D-Nucleofector™ Protocol for Jurkat clone E6.1 [ATCC®]: The Lonza Group AG provided the transfection protocol for Jurkat clone E6.1 from ATCC using 4D-Nucleofector® X Unit.

Videos on Jurkat E6.1

There are several excellent videos covering cell culture concepts, procedures, proliferation, plating, and other topics.

Passaging Cells: Cell Culture Basics: This is a great video for basic guide passaging human cell lines by ThermoFisher.
Thawing Cells: Cell Culture Basics: This video provides basic instructions on cell culturing and thawing by ThermoFisher.
Plasmid DNA Transfection Protocol: This video focused on the protocol of Plasmid DNA transfection with Lipofectamine® LTX and Plus™ Reagent, by ThermoFisher.
Measure Apoptosis using Suspension Jurkat Cells Treated with 3 Micromolar Staurosporine: This video demonstrates how to perform an apoptosis assay on the Celigo image cytometer using Hoechst and caspase 3/7 reagents by Nexcelom Bioscience.