KC-2022

293T-LRRC15 Cell Line

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Background of 293T-LRRC15 Cell Line

Leucine rich repeat containing 15 is a transmembrane protein frequently overexpressed in multiple tumor types as tumor antigen. The protein is localized to the apical plasma membrane, plasma membrane, extracellular exosome, and intracellular membrane-bounded organelles, placing it at interfaces where it can contact extracellular ligands and surrounding matrix. It also shows expression in skin, connective tissue, and the endocrine system, as well as protein expression in the gastrointestinal tract.

Specifications

Catalog Number	KC-2022
Cell Line Name	293T-LRRC15 Cell Line
NCBI/UniProt Accession Number	NM_130830.5
Clone Number	3#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous human LRRC15 gene
Quantity	One vial of frozen cells (≥2×10⁶ cells)
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	NA

Cell Line Generation

293T-LRRC15 cell line was generated using a lentiviral vector expressing the human LRRC15 sequence.

Characterization

Figure 1: Characterization of human LRRC15 overexpression in the 293T-LRRC15 stable clone using FACS.

Cell Resuscitation

Pre-warm complete culture medium (DMEM+10% FBS +1μg/mL Puromycin) 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

1. O’Prey, J., Wilkinson, S. & Ryan, K. M. Tumor Antigen LRRC15 Impedes Adenoviral Infection: Implications for Virus-Based Cancer Therapy. J. Virol. 82, 5933–5939 (2008).
2. Wang, Y. et al. LRRC15 promotes osteogenic differentiation of mesenchymal stem cells by modulating p65 cytoplasmic/nuclear translocation. Stem Cell Res. Ther. 9, 65 (2018).
3. Cui, J. et al. Expression and clinical implications of leucine-rich repeat containing 15 (LRRC15) in osteosarcoma.J. Orthop. Res. 38, 2362–2372 (2020).

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.