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	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	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

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.