KC-6193

U87MG-FAP-KO Cell Line

63949

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Background of U87MG-FAP-KO Cell Line

Fibroblast activation protein (FAP) is a type II transmembrane serine protease belonging to the dipeptidyl peptidase IV family. The protein encoded by the FAP gene is a homodimeric integral membrane gelatinase selectively expressed in reactive stromal fibroblasts of epithelial cancers, granulation tissue of healing wounds, and malignant cells of bone and soft tissue sarcomas. Under physiological conditions, FAP expression is tightly regulated and largely absent in most normal adult tissues, with transient expression observed only in some fetal mesenchymal tissues and wound healing sites. However, in over 90% of common epithelial cancers, including breast, colorectal, pancreatic, and lung carcinomas, FAP is significantly upregulated in cancer-associated fibroblasts (CAFs) within the tumor stroma. This tumor-restricted expression pattern has established FAP as an attractive diagnostic biomarker and therapeutic target. FAP contributes to tumor progression through multiple mechanisms, including extracellular matrix remodeling, promotion of cancer cell invasion and migration, and modulation of immunosuppressive tumor microenvironments. Targeting strategies currently under investigation include FAP-specific chimeric antigen receptor (CAR)-T cells, antibody-drug conjugates, and FAP-activated prodrugs, with several agents entering clinical trials.

Specifications

Catalog Number	KC-6193
Cell Line Name	U87MG-FAP-KO Cell Line
Clone Number	2A4
Host Cell Line	U87MG
Description	Stable U87MG clone with human FAP gene knockout
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	MEM+20% FBS+10% DMSO
Propagation Medium	MEM+10%FBS+0.1mM NEAA+1%Sodium pyruvate
Selection Marker	NA
Morphology	Epithelial
Subculture	Split saturated culture 1:2-1:4 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 hours
Mycoplasma Status	Negative

Cell Line Generation

U87MG-FAP-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of U87MG-FAP-KO cell line stable clone using PCR sequencing.

Figure 2: Characterization of U87MG-FAP-KO cell line stable clone using RT-PCR sequencing.

Figure 3:Characterization of U87MG-FAP-KO cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (MEM+10%FBS+0.1mM NEAA+1%Sodium pyruvate)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 1:2-1:4 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (MEM+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

Gene ID: 2191. FAP fibroblast activation protein alpha [Homo sapiens]. NCBI. Updated 2026.
Chen Z, et al. (2022). "Fibroblast activation protein-α in pancreatic adenocarcinoma: from diagnostic marker to therapeutic target." Frontiers in Oncology, 12:969731.
Mathew S, et al. (1995). "The gene for fibroblast activation protein α (FAP) maps to chromosome band 2q23." Genomics, 25(1):335-337.

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.