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The Anti-FAR (X12X217) h(28OXζ) CAR-Jurkat-NFAT-Luc cell line is a stable cell line derived from anti-FAR CAR lentivirus transduction using Jurkat-NFAT-luciferase cell as effector cell. This transduced CAR lentiviral vector was constructed to express scFv of an anti-FAR antibody linked to the CD28-OX40-CD3ζ signaling domains. The CAR-Jurkat cell line constitutively expressing the firefly luciferase gene under the control of NFAT response elements to facilitate subsequent research. And the recombinant cell product targeted FAR for CAR-T studies in human Rhabdomyosarcoma.
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Specifications
Species
Human
Host Cell Type
T lymphocyte
Overexpressed Gene
NFAT-Luciferase Reporter
Size
Containing ≥ 2 X 10*6 / vial Frozen cells
Culture Condition
Suspension
Growth Pattern
Exponential growth phase
Formulation
Frozen cells
Cell Purity
>90%
Cell Viability
>80%
Mycoplasma Testing
The cell line has been screened using the luciferase based mycoplasma detection kit to confirm the absence of mycoplasma species.
Shipping
Dry ice
Storage
Frozen cells should be stored in a liquid nitrogen tank (-150°C~-190°C).
Warnings
Avoid multiple freeze/thaw cycles
Research Use Only
Our recombinant Jurkat cells are for research use only, not for diagnostic or therapeutic use.
Quality Control
Cultures are screened for the presence of bacteries, yeast, fungi and mycoplasma (DNA amplification). Growth media are also certified based on U.S. Public Health Service Guidelines.
Tumorgenicity
Positive
Oncogenicity
Positive (In vitro life-time studies)
Sterility Testing
Creative Biolabs provides sterility testing in accordance with USP and EP regulations. All of our sterility testing is performed in an isolator or clean room environments. The cell line has been screened using the membrane filtration testing methods to confirm the absence of aerobic, anaerobic and fungi microorganisms.
Identity Testing
Identity testing is required for newly established cell lines. Isoenzyme analysis is used to confirm the identity of the species of a cell line. Alternative methods for identity testing include DNA fingerprinting, STR analysis and karyology.
Virological Safety Testing
A broad range of viruses is susceptible to affecting human cell lines. We can provide in vivo/vitro virus saftey assays by utilizing various animal systems. These viruses include: adventitious viruses, bovine viruses, human and simian viruses, porcine viruses, retrovirus and rodent viruses.
Genetic Stability Testing
We perform cell genetic stability studies under ICH guidelines. We can provide guidance on the appropriate testing program upon your requirements.
PROTOCOL
1. Thaw CAR-Jurkat cell samples quickly in a 37°C water bath until all visible ice has melted. Thaw time for a 1 ml sample in a cryovial is 2-3 minutes. Cryovials should be cool to the touch when removed from the water bath. 2. Dilute cell/medium mixture immediately with CAR-Jurkat cell culture medium. This can be performed in a single step. The dilution medium should be between 20-37°C. A dilution ratio of 1:10 (sample:medium) or greater is recommended. 3. Plate cells appropriately according to the experimental conditions of assays. 4. Culture the control lentivirus CAR-Jurkat cells or use immediately. Note: Jurkat cells that are used immediately after thawing have the highest level of viability.
Application
1. Compound screening; 2. Antibody screening; 3. Co-stimulatory and activation domain comparison; 4. Checkpoint inhibitors screening; 5. Safety switches and regulators of CAR-Jurkat functions study; 6. Pre-clinical study in vivo models; 7. CAR-Jurkat signaling, tumor microenvironment; 8. Proof of concept studies for clinical trials;
CAR Design
Target
FAR
Target Species
Human
scFv Clone
X12X217
scFv Host Spcies
Human
CAR Construction
scFv-CD28-OX40-CD3ζ
CAR Signaling Cassetes
CD28 CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatorysignals required for T cell activation and survival. CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2) proteins which are expressed on antigen-presenting cells(APC). CD28 modulates the primary TCR/CD3ζ signal in a different fashion than the late costimulatory elements OX40 and 4-1BB. CD28 enhances the expression of downstream regulators that impact on T-cell proliferation, death, differentiation, and effector functions. CAR+ T cells containing the CD28 endodomain showed strikingly enhanced sustained T cell activation, growth, survival. And CD28 results in a brightly expressed, stable receptor as the transmembrane domain. Including CD28 costimulatory domains in CARs led to enhanced anti-malignancy efficacy. OX40 OX40, also known as CD134 or TNFRSF4 is a member of the TNFR-superfamily of receptors which is not constitutively expressed on resting naïve T cells, unlike CD28. OX40 is a secondary co-stimulatory immune checkpoint molecule, expressed after 24 to 72 hours following activation. The interaction between OX40 and its binding partner, OX40L (CD252) plays an important role in antigen-specific T-cell expansion and survival. OX40 and OX40L also regulate cytokine production from T cells and modulate cytokine receptor signaling. OX40 cosignaling in CAR improve redirected T-cell effector functions and enhance anti-tumor activity. CD3ζ CD3ζ, also known as T-cell receptor zeta, which together with T-cell receptor and CD3γ, δ, ε chain, forms the TCR-CD3 complex. ζ was expressed independently from the complex. The zeta chain plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. CD3-zeta,which contains 3 ITAMs, is the most commonly used endodomain component of CARs. It transmits an activation signal to the T cell after antigen is bound. CD3-zeta may not provide a fully competent activation signal and additional co-stimulatory signaling is needed. For example, chimeric CD28 and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.
CAR Vector Name
pCDCAR1
CAR Vector length
~8kb
CAR Vector Type
Lentiviral vector
CAR Generation
Third
Targeting Diseases
Rhabdomyosarcoma
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Customer Reviews and Q&As
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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.
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