KC-4057

CHOK1-mouse-L1CAM-Cell-Line

40572

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Background of CHOK1-mouse-L1CAM-Cell-Line

L1CAM (L1 Cell Adhesion Molecule) is a Protein Coding gene.The protein encoded by this gene is an axonal glycoprotein belonging to the immunoglobulin supergene family. The ectodomain, consisting of several immunoglobulin-like domains and fibronectin-like repeats (type III), is linked via a single transmembrane sequence to a conserved cytoplasmic domain. This cell adhesion molecule plays an important role in nervous system development, including neuronal migration and differentiation. Mutations in the gene cause X-linked neurological syndromes known as CRASH (corpus callosum hypoplasia, retardation, aphasia, spastic paraplegia and hydrocephalus).

Specifications

Catalog Number	KC-4057
Cell Line Name	CHOK1-mouse-L1CAM-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous mouse L1CAM 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 + 10μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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
In Vivo Validation	NA

Cell Line Generation

CHOK1 mouse L1CAM Cell Line was generated using a lentiviral vector expressing the mouse L1CAM sequence.

Characterization

Figure 1: Characterization of mouse L1CAM overexpression in the CHO-K1 mouse L1CAM stable clone using qPCR.

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

Grońska-Pęski M, Schachner M, Hébert JM. L1cam curbs the differentiation of adult-born hippocampal neurons. Stem Cell Res. 2020 Oct;48:101999. doi: 10.1016/j.scr.2020.101999. Epub 2020 Sep 17. PMID: 32971459; PMCID: PMC7578921.
Itoh K, Fushiki S. The role of L1cam in murine corticogenesis, and the pathogenesis of hydrocephalus. Pathol Int. 2015 Feb;65(2):58-66. doi: 10.1111/pin.12245. PMID: 25641508.
Cau F, Fanni D, Manchia M, Gerosa C, Piras M, Murru R, Paribello P, Congiu T, Coni P, Pichiri G, Piludu M, Van Eyken P, Gibo Y, La Nasa G, Orrù G, Scano A, Coghe F, Saba L, Castagnola M, Faa G. Expression of L1 Cell Adhesion Molecule (L1CAM) in extracellular vesicles in the human spinal cord during development. Eur Rev Med Pharmacol Sci. 2022 Sep;26(17):6273-6282. doi: 10.26355/eurrev_202209_29651. PMID: 36111928.