KC-0848

MDA MB 468 TROP2 KO-4B1 Cell Line

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Background of MDA MB 468 TROP2 KO-4B1 Cell Line

Trop2 is expressed in normal human tissues, including breast, cervix, kidney, lung and pancreas, and is upregulated in all cancers, regardless of baseline Trop2 expression. It’s a 35-kDa transmembrane glycoprotein and calcium signal transducer encoded by the TACSTD2 gene, and is structurally related to epithelial cell adhesion molecule (EpCAM). It participates in embryonic development and is implicated in several oncogenetic signaling pathways that lead to proliferation, including the MAPK–ERK1–ERK2 pathway, nuclear factor (NF)-κB and RAF.

Specifications

Catalog Number	KC-0848
Cell Line Name	MDA MB 468 TROP2 KO-4B1 Cell Line
Host Cell Line	MDA-MB-468
Description	Stable MDA-MB-468 clone with human TROP2 gene knockout, No.4B1
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%DMEM+ 20% FBS+ 10% DMSO
Propagation Medium	DMEM + 10% FBS
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:2~1:3 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 30-40 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

MDA-MB-468-TROP2-KO-4B1 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of MDA-MB-468-TROP2-KO Cell Line stable clone using PCR sequencing.

Figure 2: Characterization of MDA-MB-468-TROP2-KO Cell Line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM + 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 1:2-1:3 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Shaffer, C. Trop2 deal heats up antibody–drug conjugate space in cancer. Nat Biotechnol 39, 128–130 (2021).