KC-1885

Raji-OS8-Cell-Line

22987

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Background of Raji-OS8-Cell-Line

Raji-OS8 stable cell was used as artificial antigen-presenting cells (APCs). OS8 could function as a membraneanchored T cell engager that directly activates TCR in T cell-based assay.

Specifications

Catalog Number	KC-1885
Cell Line Name	Raji-OS8-Cell-Line
Host Cell Line	Raji
Description	Raji cell line stable expressing exogenous membrane OKT3 scFV
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	Hygromycin B
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 OS8 cell line was generated using a lentiviral vector expressing the scFV sequence of anti-human CD3 mAb OKT3 and a C-terminal domain of mouse CD8a, which consists of transmembrane and cytoplasmic domains.

Characterization

Figure 1: Characterization of OS8 overexpressing in Raji stable clones using FACS.

Figure 2: Raji OS8 cell line was co-cultured with Jurkat-IL2P-Luc2 in the 96-well plate for 16 hours, then readout with Bright-Glo system.

Cell Resuscitation

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

Zhu, Y. et al. Identification of CD112R as a novel checkpoint for human T cells. J. Exp. Med. 213, 167ÿ176 (2016).
Wang,D. et al. Role of CD155/TIGIT in Digestive Cancers: Promising Cancer Target for Immunotherapy. Front Oncol. 2022; 12: 844260.