| Applications |
PFHR 9 (PFHR-9)is a differentiated cell line isolated as a HAT-resistant colony of the undifferentiated mouse embryonal carcinoma PCC4F in 1974. The PFHR 9 cells were not tumorigenic when injected subcutaneously into 129/Sv mice whereas the parental line did produce tumors. The cells produce basement membrane proteins - laminin-1; entactin-1; perlecan and collagen IV. The cell line is useful for the production of basement membrane proteins and as a model to study the assembly and secretion of these proteins. |
| Comments |
PFHR 9 (PFHR-9)is a differentiated cell line isolated as a HAT-resistant colony of the undifferentiated mouse embryonal carcinoma PCC4F in 1974. While PCC4F cells grow in suspension, the PRHR cell line is adherent. The cells are epithelial-like and have numerous vacuoles. PFHR 9 cells have a reported doubling time of 12 hours. The PFHR 9 cells were not tumorigenic when injected subcutaneously into 129/Sv mice whereas the parental line did produce tumors. The cells produce basement membrane proteins - laminin-1; entactin-1; perlecan and collagen IV. The cell line is useful for the production of basement membrane proteins and as a model to study the assembly and secretion of these proteins. |
| 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.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Note: Do not let the cells overgrow.
Subcultivation Ratio: 1:5 to 1:7
Medium Renewal: Every 2 to 3 days
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. |
| References |
Chung AE, et al. Morphological and biochemical observations on cells derived from the in vitro differentiation of the embryonal carcinoma cell line PCC4-F. Cancer Res. 37: 2072-2081, 1977. PubMed: 558822
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
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