KC-6249

CHOK1-ITGAM-Medium Cell Line

63037

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Background of CHOK1-ITGAM-Medium Cell Line

ITGAM (Integrin Subunit Alpha M) is a Protein Coding gene. Variations at the ITGAM gene, which encodes for the CD11b chain of the Mac-1 (alphaMbeta2; CD11b/CD18; complement receptor-3) integrin, is one of the strongest genetic risk factors for systemic lupus erythematosus (SLE). The alpha M beta 2 integrin is important in the adherence of neutrophils and monocytes to stimulated endothelium, and also in the phagocytosis of complement coated particles. Diseases associated with ITGAM include Systemic Lupus Erythematosus 6 and Neutropenia, Severe Congenital, X-Linked.

Specifications

Catalog Number	KC-6249
Cell Line Name	CHOK1-ITGAM-Medium Cell Line
NCBI/UniProt Accession Number	P11215
Clone Number	21#
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human ITGAM gene in medium level
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 +10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-ITGAM-Medium cell line was generated using a lentiviral vector expressing the human ITGAM sequence.

Characterization

Figure 1: Characterization of human ITGAM overexpression in the CHOK1-ITGAM-Medium stable clone using FACS(Kyinno, Cat#KB-1902).

Figure 2: Characterization of human ITGAM in the CHOK1-ITGAM-Medium stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (RPMI1640+10% FBS +10μg/mL Puromycin) 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, 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 in a 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 supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:4-1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

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

References

1. Yan Q, Liu Z, Chen Y, Zhang X, Zheng W, Liu X, Huang H, Liu Q, Jiang Y, Zhan S, Huang X. ITGAM-macrophage modulation as a potential strategy for treating neutrophilic Asthma: insights from bioinformatics analysis and in vivo experiments. Apoptosis. 2024 Apr;29(3-4):393-411. doi: 10.1007/s10495-023-01914-5. Epub 2023 Nov 11. PMID: 37950848.
2. Fagerholm SC, MacPherson M, James MJ, Sevier-Guy C, Lau CS. The CD11b-integrin (ITGAM) and systemic lupus erythematosus. Lupus. 2013 Jun;22(7):657-63. doi: 10.1177/0961203313491851. PMID: 23753600.
3. Ramírez-Bello J, Sun C, Valencia-Pacheco G, Singh B, Barbosa-Cobos RE, Saavedra MA, López-Villanueva RF, Nath SK. ITGAM is a risk factor to systemic lupus erythematosus and possibly a protection factor to rheumatoid arthritis in patients from Mexico. PLoS One. 2019 Nov 27;14(11):e0224543. doi: 10.1371/journal.pone.0224543. PMID: 31774828; PMCID: PMC6881022.

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.