KC-3070

CHOK1-OS8-CLEC2D-Cell-Line

43683

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Background of CHOK1-OS8-CLEC2D-Cell-Line

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, Inhibits bone resorption and Modulates the release of interferon-gamma. CLEC2D uses gene splicing to generate protein isoforms that are structurally distinct and that have different biological activities.

Specifications

Catalog Number	KC-3070
Cell Line Name	CHOK1-OS8-CLEC2D-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous OS8 CLEC2D 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS +750µg/mL Hygromycin B+10μg/ml Puromycin
Selection Marker	Puromycin,Hygromycin B
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
In Vivo Validation	NA

Cell Line Generation

CHOK1-OS8-CLEC2D Cell Line was generated using a lentiviral vector expressing the human CLEC2D sequence.

Characterization

Figure 1: Characterization of human CLEC2D overexpression in the CHO-K1 human CLEC2D stable clone using FACS sequence.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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% RPMI1640 + 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

Hu YS, Zhou H, Myers D, Quinn JM, Atkins GJ, Ly C, Gange C, Kartsogiannis V, Elliott J, Kostakis P, Zannettino AC, Cromer B, McKinstry WJ, Findlay DM, Gillespie MT, Ng KW. Isolation of a human homolog of osteoclast inhibitory lectin that inhibits the formation and function of osteoclasts. J Bone Miner Res. 2004 Jan;19(1):89-99.
Germain C, Bihl F, Zahn S, Poupon G, Dumaurier MJ, Rampanarivo HH, Padkjær SB, Spee P, Braud VM. Characterization of alternatively spliced transcript variants of CLEC2D gene. J Biol Chem. 2010 Nov 12;285(46):36207-15.