PEA 13 (ATCC® CRL-2216)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast  /  Tissue: embryo  /  Disease: LRP deficient

Organism Mus musculus, mouse
Tissue
embryo
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease LRP deficient
Age embryo; 15 days gestation
Applications
This cell line along with the MEF-1 (see ATCC CRL-2214) and PEA 10 (see ATCC CRL-2215) cell lines constitute a genetically defined system to study the endocytosis of ligands by the LRP.
These cell lines are the only available experimental system to study the effect of LRP deficiency because LRP deficient mouse embryos die in utero.
Storage Conditions liquid nitrogen vapor phase
Derivation
The line was initiated in 1993 by selection with 30 ng/ml Pseudomonas exotoxin A (PEA).
Comments
PEA 13 cells are homozygous for a disruption of the gene for low density lipoprotein receptor related protein (LRP).

Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that 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. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C

Subculture Ratio: 1:6 to 1:10
Medium Renewal: 2 to 3 times a week.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Name of Depositor TE Willnow
Year of Origin 1993
References

Kounnas MZ, et al. LDL receptor-related protein, a multifunctional ApoE receptor, binds secreted beta-amyloid precursor protein and mediates its degradation. Cell 82: 331-340, 1995. PubMed: 7543026

Willnow TE, Herz J. Genetic deficiency in low density lipoprotein receptor-related protein confers cellular resistance to Pseudomonas exotoxin A. Evidence that this protein is required for uptake and degradation of multiple ligands. J. Cell Sci. 107: 719-726, 1994. PubMed: 8006085

Orth K, et al. Low density lipoprotein receptor-related protein is necessary for the internalization of both tissue-type plasminogen activator-inhibitor complexes and free tissue-type plasminogen activator. J. Biol. Chem. 269: 21117-21122, 1994. PubMed: 8063731

Basic Documentation
References

Kounnas MZ, et al. LDL receptor-related protein, a multifunctional ApoE receptor, binds secreted beta-amyloid precursor protein and mediates its degradation. Cell 82: 331-340, 1995. PubMed: 7543026

Willnow TE, Herz J. Genetic deficiency in low density lipoprotein receptor-related protein confers cellular resistance to Pseudomonas exotoxin A. Evidence that this protein is required for uptake and degradation of multiple ligands. J. Cell Sci. 107: 719-726, 1994. PubMed: 8006085

Orth K, et al. Low density lipoprotein receptor-related protein is necessary for the internalization of both tissue-type plasminogen activator-inhibitor complexes and free tissue-type plasminogen activator. J. Biol. Chem. 269: 21117-21122, 1994. PubMed: 8063731