KC-2438

B16/F10-LY6K Cell Line

24060

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Background of B16/F10-LY6K Cell Line

Lymphocyte antigen 6 complex locus K (LY6K) is a glycosylphosphatidylinositol -anchored cell surface protein of the Ly6/uPAR family. It is overexpressed in several cancers and is investigated as a tumor-associated antigen and immunotherapy target.

Specifications

Catalog Number	KC-2438
Cell Line Name	B16/F10-LY6K Cell Line
NCBI/UniProt Accession Number	NM_017527
Clone Number	3#
Host Cell Line	B16/F10
Description	B16/F10-LY6K is a stable B16/F10 cell line overexpressing human LY6K
Quantity	One vial of frozen cells (≥2×10⁶ cells/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+1µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split at a ratio of 1:3–1:6 every 2–3 days
Incubation	37°C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Estimated 12–18 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

The B16/F10-LY6K cell line was generated using a lentiviral vector expressing the human LY6K sequence.

Characterization

Figure 1: Characterization of human LY6K expression in the B16/F10-LY6K stable clone using Western blot.

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, then 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 at 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.
Subculture the cells at a ratio of 1:3–1:6 every 2–3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% DMEM, 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 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 liquid nitrogen for long-term storage the following day.

References

Weinberg RA. The Biology of Cancer. 2nd ed. W. W. Norton & Company; 2013.
Hanahan D. Hallmarks of Cancer: New Dimensions. Cancer Discovery. 2022;12:31-46.

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.