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KC-6466

RM-1-STEAP2 Cell Line

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Home » RM-1-STEAP2 Cell Line

Background of RM-1-STEAP2 Cell Line

STEAP2 (STEAP2 Metalloreductase) is a Protein Coding gene. This gene is a member of the STEAP family and encodes a multi-pass membrane protein that localizes to the Golgi complex, the plasma membrane, and the vesicular tubular structures in the cytosol. It is highly over-expressed in various types of cancer and it up-regulation hindered cellular proliferation, invasion and metastasis abilities by inhibiting EMT process and suppressing PI3K/AKT/mTOR signaling pathway. Diseases associated with STEAP2 include Hepatic Adenomas, Familial and Prostate Cancer.

Specifications

Catalog NumberKC-6466
Cell Line NameRM-1-STEAP2 Cell Line
NCBI/UniProt Accession NumberNM_152999.3
Clone Number6#
Host Cell LineRM-1
DescriptionStable RM-1 cell line expressing exogenous STEAP2 gene
QuantityOne vial of frozen cells (≥2×106 cells)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% RPMI1640+20% FBS+10% DMSO
Propagation Medium90% RPMI1640+10% FBS
Selection MarkerNA
Morphologyfibroblast-like
SubcultureSplit saturated culture 1:4 to 1:8 every 2-3 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

Cell Line Generation

RM-1-STEAP2 cell line was generated using a lentiviral vector expressing the STEAP2 sequence.

Characterization

Figure 1:Characterization of STEAP2 in the RM-1-STEAP2 stable clone using PCR sequencing.

Figure 2. Characterization of STEAP2 overexpression in the RM-1-STEAP2 stable clone using FACS.

Cell Resuscitation

  1. Pre-warm complete culture medium (90% RPMI1640 and 10% FBS) in a 37°C water bath.
  2. Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
  3. Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
  4. Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
  5. Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
  6. Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
  7. Incubate the flask in a 37°C in a humidified 5% CO2 incubator.
  8. Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
  9. Subculture the cells at a ratio of 1:4 to 1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

  1. Prepare the freezing medium (70% RPMI1640, 20% FBS and 10% DMSO) freshly before use.
  2. Pre-chill the freezing medium on ice and label the cryovials accordingly.
  3. Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
  4. Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
  5. Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×106 cells/mL.
  6. Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
  7. Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
  8. Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

  1. Torrez CZ, Easley A, Bouamar H, Zheng G, Gu X, Yang J, Chiu YC, Chen Y, Halff GA, Cigarroa FG, Sun LZ. STEAP2 promotes hepatocellular carcinoma progression via increased copper levels and stress-activated MAP kinase activity. Sci Rep. 2024 Jun 3;14(1):12753. doi: 10.1038/s41598-024-63368-2. PMID: 38830975; PMCID: PMC11148201.
  2. Yang Q, Ji G, Li J. STEAP2 is down-regulated in breast cancer tissue and suppresses PI3K/AKT signaling and breast cancer cell invasion in vitro and in vivo. Cancer Biol Ther. 2020;21(3):278-291. doi: 10.1080/15384047.2019.1685290. Epub 2019 Nov 7. PMID: 31696760; PMCID: PMC7012168.
  3. Liu T, Niu X, Li Y, Xu Z, Chen J, Xu G. Expression and prognostic analyses of the significance of STEAP1 and STEAP2 in lung cancer. World J Surg Oncol. 2022 Mar 28;20(1):96. doi: 10.1186/s12957-022-02566-6. PMID: 35346237; PMCID: PMC8962583.

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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