KC-4262

NCI-H358-CDH3-KO-2A2-Cell-Line

43941

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Background of NCI-H358-CDH3-KO-2A2-Cell-Line

Cadherins are cell adhesion molecules that help cells to adhere to each other and help cells migrate via epithelial-mesenchymal transition (EMT) . They control calcium-dependent cellular adhesion and bind to various cell types and the extracellular matrix (ECM), maintaining cellular structure.Cadherin 3 (CDH3, also known as PCAD) has a molecular weight of 118kDa; comprises extracellular, transmembrane, and cytoplasmic domains; and promotes homotypic interactions. This gene encodes a classical cadherin of the cadherin superfamily. Alternative splicing results in multiple transcript variants, at least one of which encodes a preproprotein that is proteolytically processed to generate the mature glycoprotein.In normal cells, CDH3 regulates cell growth and migration, but CDH3 may have opposing effects in tumors depending on type: CDH3 promotes tumorigenesis in pancreatic, breast, and gastric cancer but has the opposite effect in non-small cell lung carcinoma, liver cancer, and melanoma .

Specifications

Catalog Number	KC-4262
Cell Line Name	NCI-H358-CDH3-KO-2A2-Cell-Line
Host Cell Line	NCI-H358
Description	Stable NCI-H358 clone with human CDH3 gene knockout, No.2A2
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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:6 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 38 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

NCI-H358-CDH3-KO-2A2 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of NCI-H358-CDH3-KO-2A2 cell line stable clone using PCR sequencing.

Figure 2: Characterization of NCI-H358-CDH3-KO-2A2 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of NCI-H358-CDH3-KO-2A2 cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS)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 1:4-1:6 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

Sharma G, Mo JS, Lamichhane S, Chae SC. MicroRNA 133A Regulates Cell Proliferation, Cell Migration, and Apoptosis in Colorectal Cancer by Suppressing CDH3 Expression. J Cancer. 2023 Apr 9;14(6):881-894. doi: 10.7150/jca.82916. PMID: 37151391; PMCID: PMC10158507.
https://www.ncbi.nlm.nih.gov/gene/1001