KC-0412

MKN45-Cell-Line-(Not-for-sale)

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Home » MKN45-Cell-Line-(Not-for-sale)

Background of MKN45-Cell-Line-(Not-for-sale)

MKN45 was established from the poorly differentiated adenocarcinoma of the stomach (medullary type) of a 62-year-old woman

Specifications

Catalog NumberKC-0412
Cell Line NameMKN45-Cell-Line-(Not-for-sale)
Host Cell LineNA
DescriptionHuman gastric cancer cell line, established from the poorly differentiated adenocarcinoma of the stomach (medullary type) of a 62-year-old woman
QuantityTwo vials of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
Application3D cell culture; Cancer research
Freezing Medium70% RPMI1640+20% FBS+10% DMSO
Propagation MediumRPMI1640+10%FBS
Selection MarkerNA
MorphologyBoth spindle-shaped or oval cells growing in monolayers and single round cells or clumps in suspension
SubcultureSplit saturated culture 1:3-1:5 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 30-38 hours
Mycoplasma StatusNegative
In Vivo ValidationYes

Cell Line Generation

NA

Characterization

Cell Resuscitation

  1. Prewarm culture medium (RPMI1640 + 10% FBS)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 1:3-1:5 every 2-3 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. Forty-nine gastric cancer cell lines with integrative genomic profiling for development of c-MET inhibitor.
  2. Int. J. Cancer 143:151-159(2018)
  3. Comparison between the gastric cancer cell line MKN-45 and the high-potential peritoneal dissemination gastric cancer cell line MKN-45P.
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