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KC-1906

PC3-PDL1-Trop2-Cell-Line

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Datasheet
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Home » 细胞系 » PC3-PDL1-Trop2-Cell-Line

Background of PC3-PDL1-Trop2-Cell-Line

PD-L1, also called human programmed cell death ligand 1, is a transmembrane protein that plays a major role in suppressing the immune system during particular events such as pregnancy, tissue allografts, autoimmune disease, virus infection, and cancer. PD-L1 binds to its receptor PD-1 on activated T cells, B cells, and myeloid cells, to modulate activation or inhibition. Upregulation of PD-L1 can allow the cancer cell to evade the host immune system. Trop2, also named as epithelial glycoprotein 1 antigen (EGP-1), is a member of a family including at least two type I membrane protein, Trop2 acts as a cell surface receptor and transduce intracellular calcium signal, and is also a carcinoma associated antigen, which was the target of several therapeutic antibodies, including GA773, and sacituzumab govitecan.

Specifications

Catalog NumberKC-1906
Cell Line NamePC3-PDL1-Trop2-Cell-Line
Host Cell LineHuman PC3 cell line
DescriptionPC3 cell line stable expressing exogenous human PDL1 and TROP2 gene
QuantityOne vial of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% F12K+20% FBS+10% DMSO
Propagation MediumHam's F12K+10%FBS+1µg/mL Puromycin+100µg/mL Hygromycin B
Selection MarkerPuromycin, HygromycinB
MorphologyEpithelial
SubcultureSplit saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 24 hours
Mycoplasma StatusNegative
In Vivo ValidationNA

Cell Line Generation

PC3 Human PDL1-TROP2 Cell Line was generated using a lentiviral plasmid with human TROP2 gene sequence.

Characterization

Cell Resuscitation

  1. Prewarm culture medium (Ham's F12K + 10% FBS + 1µg/mL Puromycin + 100µg/mL Hygromycin B) 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 2-3 days; seed out at about 1-2 × 105 cells/mL.

Cell Freezing

  1. Prepare the freezing medium (70% F12K + 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. Sunshine, J. & Taube, J. M. PD-1/PD-L1 inhibitors. Current Opinion in Pharmacology 23, 32ÿ38 (2015).
  2. Boussiotis, V. A. Molecular and Biochemical Aspects of the PD-1 Checkpoint Pathway. N Engl J Med 375, 1767ÿ 1778 (2016).
  3. Topalian, S. L. et al. Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N Engl J Med 366, 2443ÿ2454 (2012).
  4. Linnenbach AJ, Wojcierowski J, Wu SA, et al. (1989). "Sequence investigation of the major gastrointestinal tumor-associated antigen gene family, GA733". Proc. Natl. Acad. Sci. U.S.A. 86 (1): 27ÿ31.
  5. Fornaro M, Dell'Arciprete R, Stella M, et al. (1995). "Cloning of the gene encoding Trop-2, a cell-surface glycoprotein expressed by human carcinomas". Int. J. Cancer. 62 (5): 610ÿ8.
  6. El Sewedy T, Fornaro M, Alberti S (1998). "Cloning of the murine TROP2 gene: conservation of a PIP2-binding sequence in the cytoplasmic domain of TROP-2". Int. J. Cancer. 75 (2): 324ÿ30.

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.

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