FAP, also known as FAPalpha, is a type II transmembrane glycoprotein that belongs to the serine protease family, which is selectively expressed in reactive stromal fibroblasts of epithelial cancers, granulation tissue of healing wounds, and malignant cells of bone and soft tissue sarcomas. FAP is thought to be involved in the control of fibroblast growth or epithelial-mesenchymal interactions during development, tissue repair, and epithelial carcinogenesis. Acts as a tumor suppressor in melanocytic cells through regulation of cell proliferation and survival in a serine protease activity-independent manner.
Specifications
Catalog Number:
KC-2635
Cell Line Name:
HT1080 FAP Cell Line
Price:
0
Host Cell Line:
Human HT1080 cell line
Description:
Stable HT1080 clone expressing exogenous human FAP
Quantity:
One vial of frozen cells (5X106 per vial)
Stability:
Stable in culture over a minimum of 10 passages
Application:
Drug screening and biological assays
Freeze Medium:
70% RPMI1640+20% FBS+10% DMSO
Propagation Medium:
RPMI1640+10%FBS+1µg/mL Puromycin
Selection Marker:
Puromycin
Morphology:
Epithelial
Subculture:
Split saturated culture 1:4-1:8 every 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 48 hours
Mycoplasma Status:
Negative
Cell Line Generation
HT-1080 human FAP cell line was generated using a lentiviral vector expressing the human FAP sequence.
Characterization using PCR sequencing and WB
0
Application
0
Cell Resuscitation
1. Prewarm culture medium (RPMI1640 + 10% FBS + 1µg/mL Puromycin) in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:4-1:8 every 3 days; seed out at about 1-3 × 105 cells/mL.
Cell Freezing
1. Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage
References
1. "Entrez Gene: fibroblast activation protein, alpha".
2. Puré E, Blomberg R (August 2018). "Pro-tumorigenic roles of fibroblast activation protein in cancer: back to the basics". Oncogene. 37 (32): 4343–4357. doi:10.1038/s41388-018-0275-3. PMC 6092565. PMID 29720723.
3. Busek P, Mateu R, Zubal M, Kotackova L, Sedo A (June 2018). "Targeting fibroblast activation protein in cancer - Prospects and caveats". Frontiers in Bioscience. 23: 1933–1968. doi:10.2741/4682. PMID 29772538.
4. Niedermeyer J, Garin-Chesa P, Kriz M, Hilberg F, Mueller E, Bamberger U, Rettig WJ, Schnapp A (April 2001). "Expression of the fibroblast activation protein during mouse embryo development". The International Journal of Developmental Biology. 45 (2): 445–7. PMID 11330865.
5. Dolznig H, Schweifer N, Puri C, Kraut N, Rettig WJ, Kerjaschki D, Garin-Chesa P (August 2005). "Characterization of cancer stroma markers: in silico analysis of an mRNA expression database for fibroblast activation protein and endosialin". Cancer Immunity. 5: 10. PMID 16076089.
