KC-1835

Raji-OS8-PVR-Cell-Line

22895

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Background of Raji-OS8-PVR-Cell-Line

The poliovirus receptor PVR, also known as CD155 or NECL-5. PVR mediates natural killer cell adhesion and triggers its effector function, participates in mediating tumor cell infiltration and migration, and is the attachment receptor of poliovirus. PVR is expressed in monocytes, macrophages, thymocytes and neurons of the central nervous system. PVR is able to bind CD226, DNAX accessory molecule-1 (DNAM-1), T Cell-Activated Increased Late Expression Protein, TACTILE (CD96), and T Cell Immunoreceptor with Ig and ITIM domains (TIGIT). PVR expression was associated with an unfavorable prognosis in solid tumors such as colon cancer, breast cancer, lung adenocarcinoma, pancreatic cancer, melanoma, and glioblastoma. Nectin Therapeutics is developing a CD155 antibody, NTX-1088, in clinical phase. Raji-OS8 stable cell was used as an artificial antigen presenting cells (APCs), OS8 could function as a membrane anchored T cell engager that directly activates TCR in T cell-based assay.

Specifications

Catalog Number	KC-1835
Cell Line Name	Raji-OS8-PVR-Cell-Line
Host Cell Line	Human Raji cell line
Description	Stable Raji cell line expressing exogenous human PVR gene
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+400µg/mL Hygromycin B
Selection Marker	HygromycinB
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Raji human OS8 PVR cell line was generated using a lentiviral vector expressing the human OS8 PVR sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 400µg/mL Hygromycin B)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Koike S, Horie H, Ise I, Okitsu A, Yoshida M, Iizuka N, Takeuchi K, Takegami T, Nomoto A (October 1990). "The poliovirus receptor protein is produced both as membrane-bound and secreted forms". EMBO J. 9 (10): 3217ÿ 24.
Mendelsohn CL, Wimmer E, Racaniello VR (1989). "Cellular receptor for poliovirus: molecular cloning, nucleotide sequence, and expression of a new member of the immunoglobulin superfamily". Cell. 56 (5): 855ÿ 65.
Maier MK, Seth S, Czeloth N, et al. (2007). "The adhesion receptor CD155 determines the magnitude of humoral immune responses against orally ingested antigens". European Journal of Immunology. 37 (8): 2214ÿ25.