PNEC30 (ATCC® CRL-2930)

Organism: Mus musculus, transgenic, mouse, transgenic  /  Cell Type: neuroendocrine  /  Tissue: prostate  /  Disease: prostate neuroendocrine cancer

Organism Mus musculus, transgenic, mouse, transgenic
Tissue prostate
Cell Type neuroendocrine
Product Format frozen
Morphology neuronal
Culture Properties adherent
Biosafety Level 1
Disease prostate neuroendocrine cancer
Age 6 months
Gender male
Applications
PNEC cell lines should be useful for genetic and/or pharmacologic studies of the regulation of NE cell proliferation, differentiation, and tumorigenesis.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
Prostate neuroendocrine cancer (PNEC) cell lines were established from CR2-TAg prostate tumors and metastases.
Clinical Data
6 months
male
Receptor Expression
Gamma-aminobutyric acid (GABAA), expressed
Genes Expressed
mAsh1 transcription factor, expressed
Cellular Products
mAsh1 transcription factor, expressed
Tumorigenic YES RefHu Y, et al. RNA interference of achaete-scute homolog 1 in mouse prostate neuroendocrine cells reveals its gene targets and DNA binding sites. Proc. Natl. Acad. Sci. 101(15):5559-5564, 2004. PubMed: 15060276
Effects
tumorigenic in BALB/c mice
Comments

GeneChip analyses of cell lines harvested at different passages, and as xenografted tumors, indicated that PNECs express consistent features ex vivo and in vivo and share a remarkable degree of similarity with primary CR2-TAg prostate neuroendocrine (NE) tumors. 

PNECs express mAsh1 (mouse homolog proneural gene complex achaete-scute), a basic helix-loop-helix (bHLH) transcription factor essential for NE cell differentiation in other tissues. PNEC cell lines should be useful for genetic and/or pharmacologic studies of the regulation of NE cell proliferation, differentiation, and tumorigenesis.

PNEC30 cells, when cultured on non-coated surfaces, grow in suspension as multicellular aggregates that resemble the neurospheres of cultured neural stem cells.
Complete Growth Medium The base medium for this cell line is Neural Progenitor Basal Medium, which is supplied as part of the NPMM Neural Progenitor Maintenance Medium Bullet Kit available from Lonza/Clonetics Inc., Catalog No. CC-3209. To make the complete growth medium, add the following components to 500 ml of the base medium:
  • additives that are supplied with the kit (ATCC does not use gentamycin-amphotericin B)
  • heat-inactivated fetal bovine serum (FBS) to a final concentration of 10%
  • bovine pituitary extract (BPE) (Lonza/Clonetics, Inc., Catalog No. CC-4009) to a final concentration of 0.3%

Note: Do not filter complete medium.
Subculturing

Volumes used in this protocol are for 75 cm2. flasks; proportionally reduce or increase amount of solutions for culture vessels of other sizes.


Note: These cells are cultured on poly-D-lysine coated vessels (BD BioCoat Cellware, BD Biosciences, Cat. No. 356524 for 75 cm2. flask) which are additionally coated with 20 μg/mL laminin (Sigma, Cat. No. L2020 or equivalent). Add 5 mL laminin solution to a 75 cm2. flask and incubate overnight at room temperature. Remove laminin solution and allow flask to air dry uncapped and standing upright in a biological cabinet before introducing cells. 
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new poly-D-lysine and laminin coated culture vessels. An inoculum of 2 X 104 to 5 X 104 viable cells/cm2 is recommended.
  7. Incubate cultures at 37°C. subculture when cultures reach a cell concentration between 1 X 105 and 2 X 105 cells/cm2

Subcultivation ratio: A subcultivation ratio of 1:3 to 1:8 is recommended.
Medium renewal: Every 2 to 3 days.

Cryopreservation
Freeze medium: fetal bovine serum, 90%; DMSO, 10%
liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Population Doubling Time approximately 50 hours
Name of Depositor J Gordon and J Ippolito
Year of Origin 2002
References

Ippolito JE, et al. An integrated functional genomics and metabolomics approach for defining poor prognosis in human neuroendocrine cancers. Proc. Natl. Acad. Sci. 102(28):9901-9906, 2005. PubMed: 15998737

Hu Y, et al. RNA interference of achaete-scute homolog 1 in mouse prostate neuroendocrine cells reveals its gene targets and DNA binding sites. Proc. Natl. Acad. Sci. 101(15):5559-5564, 2004. PubMed: 15060276

Hu Y, et al. RNA interference of achaete-scute homolog 1 in mouse prostate neuroendocrine cells reveals its gene targets and DNA binding sites. Proc. Natl. Acad. Sci. 101(15):5559-5564, 2004. PubMed: 15060276

Basic Documentation
Other Documentation
References

Ippolito JE, et al. An integrated functional genomics and metabolomics approach for defining poor prognosis in human neuroendocrine cancers. Proc. Natl. Acad. Sci. 102(28):9901-9906, 2005. PubMed: 15998737

Hu Y, et al. RNA interference of achaete-scute homolog 1 in mouse prostate neuroendocrine cells reveals its gene targets and DNA binding sites. Proc. Natl. Acad. Sci. 101(15):5559-5564, 2004. PubMed: 15060276

Hu Y, et al. RNA interference of achaete-scute homolog 1 in mouse prostate neuroendocrine cells reveals its gene targets and DNA binding sites. Proc. Natl. Acad. Sci. 101(15):5559-5564, 2004. PubMed: 15060276