KC-6730

293T-FCRL5 Cell Line

66239

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Background of 293T-FCRL5 Cell Line

FCRL5 (Fc Receptor Like 5) , also known as CD307e, FCRH5, IRTA2, or BXMAS1, is a single-pass type I membrane protein belonging to the immunoglobulin receptor superfamily. It is encoded by a gene located on chromosome 1q23.1 and is primarily expressed in lymphoid tissues such as lymph node and spleen. FCRL5 functions as a dual regulator of B cell receptor (BCR) signaling and is implicated in B-cell development and lymphomagenesis. In multiple myeloma (MM), FCRL5 is considerably upregulated and has emerged as a promising immunotherapeutic target. Therapeutic strategies targeting FCRL5 include CAR-T cells, bispecific T-cell engagers (e.g., Cevostamab), and antibody-drug conjugates (ADCs).

Specifications

Catalog Number	KC-6730
Cell Line Name	293T-FCRL5 Cell Line
NCBI/UniProt Accession Number	NM_031281.3
Clone Number	8#
Host Cell Line	293T cell line
Description	Stable 293T cell line expressing exogenous human FCRL5 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% basal medium+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS+1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	N/A

Cell Line Generation

The 293T-FCRL5 cell line was generated using a lentiviral vector expressing the human FCRL5 sequence.

Characterization

Cell Resuscitation

Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:4-1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.