KC-1933-DW

MC38-LRRC15 Cell Line

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Background of MC38-LRRC15 Cell Line

LRRC15 (Leucine Rich Repeat Containing 15) is a transmembrane protein that enables several functions, including collagen binding activity, integrin binding activity, and TGF-β sequestration. It is involved in several biological processes, such as negative regulation of TGF-β signaling pathway, modulation of immune cell infiltration, and tissue fibrosis. Located primarily in the plasma membrane, LRRC15 is highly active at the cell surface where it interacts with extracellular matrix components. Its specific expression on cancer-associated fibroblasts (CAFs) in multiple solid tumors positions LRRC15 as a key regulator of the tumor microenvironment and an emerging therapeutic target for modulating stromal biology in oncology.

Specifications

Catalog Number	KC-1933-DW
Cell Line Name	MC38-LRRC15 Cell Line
Clone Number	4#
Host Cell Line	MC38
Description	Stable MC38 cell line expressing exogenous LRRC15 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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS +5μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

MC38-LRRC15 cell line was generated using lentivirus expressing LRRC15 sequence.

Characterization

Figure 1. Characterization of LRRC15 over-expression in the MC38-LRRC15 stable clone using FACS.[Primary antibody: PR1498487-G1-LRRC15, Cat#KA-1254, Kyinno]

Figure 2:Validation of in vivo tumorigenicity of MC38 cells stably expressing LRRC15 via subcutaneous implantation in C57BL/6 mice, with tumor growth monitored by measuring volume (V=0.5×length×width²) and body weight.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS, 5μg/mL Puromycin)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% DMEM + 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

Tang H, Liu W, Xu Z, Zhao J, Wang W, Yu Z, Wei M. Integrated microenvironment-associated genomic profiles identify LRRC15 mediating recurrent glioblastoma-associated macrophages infiltration. J Cell Mol Med. 2021 Jun;25(12):5534-5546. doi: 10.1111/jcmm.16563. Epub 2021 May 7. PMID: 33960636; PMCID: PMC8184692.
Purcell JW, Tanlimco SG, Hickson J, Fox M, Sho M, Durkin L, Uziel T, Powers R, Foster K, McGonigal T, Kumar S, Samayoa J, Zhang D, Palma JP, Mishra S, Hollenbaugh D, Gish K, Morgan-Lappe SE, Hsi ED, Chao DT. LRRC15 Is a Novel Mesenchymal Protein and Stromal Target for Antibody-Drug Conjugates. Cancer Res. 2018 Jul 15;78(14):4059-4072. doi: 10.1158/0008-5472.CAN-18-0327. Epub 2018 May 15. PMID: 29764866.