KC-5309

Raji-LLT1 Cell Line

55654

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Background of Raji-LLT1 Cell Line

LLT1 aliase is CLEC2D. CLEC2D (C-Type Lectin Domain Family 2 Member D) is a Protein Coding gene. This gene encodes a member of the natural killer cell receptor C-type lectin family. The encoded protein inhibits osteoclast formation and contains a transmembrane domain near the N-terminus as well as the C-type lectin-like extracellular domain. Several alternatively spliced transcript variants have been identified for this gene.

Specifications

Catalog Number	KC-5309
Cell Line Name	Raji-LLT1 Cell Line
Clone Number	5#
Host Cell Line	Raji
Description	Raji cell line stably expressing exogenous human LLT1 gene
Quantity	One vial 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 + 2 μg/mL Puromycin
Selection Marker	Puromycin
Morphology	lymphoblast
Subculture	Split saturated culture 1:6-1:8 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

Cell Line Generation

Raji-LLT1 cell line was generated using a lentiviral vector expressing the human LLT1 sequence

Characterization

Figure1: Characterization of LLT1 overexpressing in Raji stable clones using FACS.

Figure2: Characterization of Raji-LLT1 cell line stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 2 μg/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 1:6-1:8 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

de Vicente JC, Lequerica-Fernández P, Rodrigo JP, Rodríguez-Santamarta T, Blanco-Lorenzo V, Prieto-Fernández L, Corte-Torres D, Vallina A, Domínguez-Iglesias F, Álvarez-Teijeiro S, García-Pedrero JM. Lectin-like Transcript-1 (LLT1) Expression in Oral Squamous Cell Carcinomas: Prognostic Significance and Relationship with the Tumor Immune Microenvironment. Int J Mol Sci. 2024 Apr 13;25(8):4314. doi: 10.3390/ijms25084314. PMID: 38673902; PMCID: PMC11050533.
Boles KS, Barten R, Kumaresan PR, Trowsdale J, Mathew PA. Cloning of a new lectin-like receptor expressed on human NK cells. Immunogenetics. 1999 Oct;50(1-2):1-7. doi: 10.1007/s002510050679. PMID: 10541800.

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.