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Nerve growth factor receptor (Ngfr); Protein and mRNA, negative RefHai M, et al. Comparative analysis of Schwann cell lines as model systems for myelin gene transcription studies. J. Neurosci. Res. 69: 497-508, 2002. PubMed: 12210843

Platelet derived growth factor receptor, alpha; negative RefBadache A, De Vries GH. Neurofibrosarcoma-derived Schwann cells overexpress platelet-derived growth factor (PDGF) receptors and are induced to proliferate by PDGF BB. J. Cell. Physiol. 177: 334-342, 1998. PubMed: 9766530

Platelet derived growth factor receptor, beta; negative RefBadache A, De Vries GH. Neurofibrosarcoma-derived Schwann cells overexpress platelet-derived growth factor (PDGF) receptors and are induced to proliferate by PDGF BB. J. Cell. Physiol. 177: 334-342, 1998. PubMed: 9766530

cyclic nucleotide phosphodiesterase 1 (Cnp1) (CNPase); positive laminin; positive RefBadache A, De Vries GH. Neurofibrosarcoma-derived Schwann cells overexpress platelet-derived growth factor (PDGF) receptors and are induced to proliferate by PDGF BB. J. Cell. Physiol. 177: 334-342, 1998. PubMed: 9766530  

Myelin-associated glycoprotein (Mag); Protein and mRNA, negative. Myelin basic protein (Mbp); Protein and mRNA, negative. Myelin protein zero (Mpz) (Charcot-Marie-Tooth neuropathy 1B); Protein and mRNA, negative. Peripheral myelin protein 22 (Pmp22); mRNA, positive; protein, negative. S100 calcium-binding protein, beta (neural) (S100b) protein; positive RefHai M, et al. Comparative analysis of Schwann cell lines as model systems for myelin gene transcription studies. J. Neurosci. Res. 69: 497-508, 2002. PubMed: 12210843

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-053mM 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. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:6 to 1:10
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.

Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO₂), 5%

Hai M, et al. Comparative analysis of Schwann cell lines as model systems for myelin gene transcription studies. J. Neurosci. Res. 69: 497-508, 2002. PubMed: 12210843

Badache A, De Vries GH. Neurofibrosarcoma-derived Schwann cells overexpress platelet-derived growth factor (PDGF) receptors and are induced to proliferate by PDGF BB. J. Cell. Physiol. 177: 334-342, 1998. PubMed: 9766530

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.