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Anti-CEA (MFE23)-FcRɣ+PI3K CAR THP1 (CARTHP-W0234)

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All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

The anti-CEA chimeric antigen receptor (CAR) THP1 cell line is a stable cell line developed using anti-CEA CAR lentivirus transfection. This transfected CAR lentiviral vector is constructed to express a scfv of anti-CEA antibody (MFE23) linked to FcRɣ+PI3K signaling domains.The anti-CEA CAR engineered THP1 cells can be used to be differentiated into macrophage cells for in vitro or in vivo phagocytosis studies. This CAR-THP1 cell line product can be used for the researches of Ovarian, gastrointestinal, colorectal, hepatocellular carcinoma (HCC).

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  • Marker:
  • Positive rate:
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Details

  • Target
  • CEA
  • Cell Type
  • THP1
  • Cell Background
  • THP-1 is a human leukemia monocytic cell line, which has been extensively used to study monocyte/macrophage functions, mechanisms, signaling pathways, and nutrient and drug transport. This cell line has become a common model to estimate modulation of monocyte and macrophage activities. CAR engineered THP1 cells can be differentiated into macrophages using PMA (Phorbol 12-myristate 13-acetate). Once differentiated (M0 macrophages), they were incubated with IL-4 and IL-13 in order to obtain M2 polarized macrophages or with IFN-gamma and LPS for classical macrophage activation (M1).
  • Targeting Diseases
  • Ovarian, gastrointestinal, colorectal, hepatocellular carcinoma (HCC)
  • CAR Expression
  • Positive
  • CAR Positive Rate
  • >95% for stable cell pool (please submit an inquiry if single clonal cell line is needed.)
  • Cell Viability
  • >90%
  • Mycoplasma Testing
  • Negative
  • Handling Notes
  • Frozen cells should be thawed immediately upon receipt and grown according to handling procedure to ensure cell viability and proper assay performance.
    Note: Do not freeze the cells upon receipt as it may result in irreversible damage to the cell line.
    Disclaimer: We cannot guarantee cell viability if the cells are not thawed immediately upon receipt and grown according to handling procedure.
  • Safety Consideration
  • Biosafety Level 1
  • Restriction
  • Research use only
  • Size
  • 2 million cells*2 vials
  • Storage
  • Frozen cells should be stored in a liquid nitrogen tank (-150°C~-190°C) for long term.

CAR Components

  • Promoter
  • EF1a
  • scFv
  • MFE23
  • Hinge
  • CD8
  • TM
  • CD8
  • ICD
  • FcRɣ+PI3K
  • Marker
  • GFP or N/A
  • Receptor Construction
  • scFv-hinge(CD8)-TM(CD8)-ICD(FcRɣ+PI3K)
  • Discription of Signaling Cassetes
  • FcRɣ
    FcRɣ is used as the CAR intracellular signaling domain. As a phagocytosis receptor, FcRɣ has cytosolic Immunoreceptor Tyrosine-based Activation Motifs (ITAMs) that are phosphorylated by Src family kinases. Based on this mechanism, FcRɣ signaling can induce macrophage phagocytosis which has been demonstrated in the published article.
    Fc gamma receptors belong to the group of non-catalytic tyrosine-phosphorylated receptors which share a similar signalling pathway involving phosphorylation of tyrosine residues. The receptors generate signals within their cells through an important activation motif known as an immunoreceptor tyrosine-based activation motif (ITAM). An ITAM is present in the intracellular tail of FcγRIIA, and its phosphorylation induces phagocytosis in macrophages. FcγRI and FcγRIIIA do not have an ITAM but can transmit an activating signal to their phagocytes by interacting with another protein that does. This adaptor protein is called the Fcγ subunit and, like FcγRIIA, contains the two YXXL sequences that are characteristic of an ITAM.

    PI3K
    Previous work demonstrated that PI3K signaling is important for internalization of large targets. To increase PI3K recruitment to the CAR-P, we fused the portion of the CD19 cytoplasmic domain (amino acids 500 to 534) that recruits the p85 subunit of PI3K to the CAR. A CAR-P containing the p85 recruitment motif alone (CAR-P-PI3K) was able to induce some whole cell engulfment. Assembling an array of motifs designed to recruit distinct phagocytic effectors can increase CAR-P activity.

Target

  • Target
  • CEA
  • Target Species
  • Human
  • Clone
  • MFE23
  • Host
  • Mouse
  • Gene Name
  • CEA cell adhesion molecule 5
  • Introduction
  • This gene encodes a cell surface glycoprotein that represents the founding member of the carcinoembryonic antigen (CEA) family of proteins. The encoded protein is used as a clinical biomarker for gastrointestinal cancers and may promote tumor development through its role as a cell adhesion molecule. Additionally, the encoded protein may regulate differentiation, apoptosis, and cell polarity. This gene is present in a CEA family gene cluster on chromosome 19. Alternative splicing results in multiple transcript variants.
  • Alternative Names
  • CEA; CD66e; CEA cell adhesion molecule 5; CEACAM5

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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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