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Application: Use as positive control in the development of anti-GPC3 CAR-T cells.Screen modulators of anti-GPC3 CAR-T cytotoxicity.Design and optimize co-culture cytotoxicity assays for anti-GPC3 specific CAR-T cell evaluation.
Background: GPC3, also known as Glypican-3 and OCI5, belongs to the glypican family and is highly expressed in the lungs, liver and kidneys. Its function is tissue dependent and can either promote or suppress tumorigenesis. Being a heparin sulfate proteoglycan, it is overexpressed in neoplasms including malignant melanoma, hepatocellular carcinoma, and testicular yolk sac tumors and plays a significant role in cell growth and differentiation. Due to its highly restricted expression in normal tissues and high prevalence in many solid tumors, GPC3 has become an attractive target for chimeric antigen receptor (CAR) T cell therapy.
Description: Anti-GPC3 CAR-T Cells are produced by high-titer lentiviral transduction of human primary CD4+ and CD8+ T cells using Anti-GPC3 CAR Lentivirus (Clone GC33-CD28TM-41BB-CD3ζ) (BPS Bioscience #82494). These ready-to-use CAR-T cells express an anti-GPC3 (glypican-3) CAR (chimeric antigen receptor) consisting of the ScFv portion of anti-GPC3 (clone GC33) linked to a 2nd generation CAR, containing IgG1 short hinge and CD28 transmembrane domains, 4-1BB and CD3ζ signaling domains (Figure 1).These CAR-T cells have been validated using flow cytometry (to determine CAR expression) and co-culture cytotoxicity assays. Figure 1. Construct diagram showing components of the Anti-GPC3 CAR expressed in Anti-GPC3 CAR-T Cells.
Mycoplasma Testing: The cells have been screened to confirm the absence of Mycoplasma species.
Storage Stability: Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage.
Supplied As: Each vial contains 2 x 106 cells in 1 ml of CryoStor® CS10 (Stemcell Technologies #100-1061)
Target: GPC3
Uniprot: P51654
Unit Definition: Primary Cells
Warnings: Avoid freeze/thaw cycles.Donors have been screened and determined negative for:Hepatitis B (anti-HBc EIA, HBsAg EIA)Hepatitis C (anti-HCV EIA)Human Immunodeficiency Virus (HIV-1/HIV-2 plus O)Human T-Lymphotropic Virus (HTLV-I/II)HIV-1/HCV/HBVWest Nile VirusTrypanasoma cruziNote: Testing cannot guarantee that any sample is completely virus-free. These cells should be treated as potentially infectious and appropriate Biological Safety Level 2 (BSL-2) precautions should be used.
Biosafety Level: BSL-1
References: Li W., et al., 2017 Hum Gene Ther. 28(5):437-448.
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