KC-2397

RM1-FOLH-Luc2-Cell-Line

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Home » 细胞系 » RM1-FOLH-Luc2-Cell-Line

Background of RM1-FOLH-Luc2-Cell-Line

FOLH, also known as PSMA, is a type II transmembrane glycoprotein belonging to the M28 peptidase family, which is a glutamate-preferrng carboxy-peptidase. PSMA is highly expressed in prostate secretory-acinar epithelium, in some benign extraprostatic epithelia cellsfrom breast, duodenum, and kidney tissues, and in prostate cancer. In the prostate the protein is up-requlated in cancerous cells and is used as an effective diagnostic and prognostic indicator of prostate cancer.

Specifications

Catalog NumberKC-2397
Cell Line NameRM1-FOLH-Luc2-Cell-Line
Host Cell LineMouse RM1 cell line
DescriptionStable RM1 cell line expressing exogenous FOLH gene
QuantityTwo vials of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% RPMI1640+20% FBS+10% DMSO
Propagation MediumRPMI1640+10%FBS+2μg/mL Puromycin
Selection MarkerPuromycin
MorphologyFibroblast-like
SubcultureSplit saturated culture 1:3-1:6 every 3 days; seed out at about 1-3 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative
In Vivo ValidationYes

Cell Line Generation

RM1 Human FOLH Luc2 cell line was generated using a lentiviral vector expressing the human FOLH sequence.

Characterization

Cell Resuscitation

  1. Prewarm culture medium (RPMI1640 + 10% FBS + 2μg/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 1:3-1:6 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. Prostate-Specific Membrane Antigen (PSMA) Theranostics for Treatment of Oligometastatic Prostate Cancer.PMID: 34829977 PMCID: PMC8621856 DOI: 10.3390/ijms222212095
  2. GCPII imaging and cancer. C A Foss 1, R C Mease, S Y Cho, H J Kim, M G Pomper.PMID: 22304713 PMCID: PMC4076792 DOI: 10.2174/092986712799462612
  3. Is prostate-specific membrane antigen a multifunctional protein Ayyappan? K Rajasekaran 1, Gopalakrishnapillai Anilkumar, Jason J Christiansen.PMID: 15840561 DOI: 10.1152/ajpcell.00506.2004
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