Mouse spinal cord neurons in serum-free culture media: suitability for patch clamp studies on chemical and electrical excitability.
Methods were devised for the serum-free culture of spinal cord neurons derived from 12- to 13-day mouse embryos. Neurons exhibited good attachment if plated for 24 h on poly-d-lysine-coated dishes in the presence of serum. Cultures were subsequently fed with a serum-free medium consisting of minimum essential medium, Earle's salts and the N1 supplement, i.e. insulin, putrescine, transferrin, progesterone and selenium. After 3 weeks in vitro, growth and survival of neurons in this medium were comparable to results obtained using serum-supplemented medium. The presence of putrescine was not essential for the beneficial effects of N1, while insulin was required for long-term survival in serum-free media. Neurons maintained in serum-free media for 3 weeks retained aspects of electrical and chemical excitability characteristic of serum-grown cells.
Salamanca
M C
MC
Department of Physiology, University of British Columbia, Vancouver, Canada.
Mathers
D A
DA
D000818:Animals; D003470:Culture Media; D003584:Cytological Techniques; D004594:Electrophysiology; D007328:Insulin / Q000494:pharmacology; D051379:Mice / Q000196:embryology; D008815:Mice, Inbred Strains; D009462:Neurology / Q000379:methods*; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D011700:Putrescine / Q000494:pharmacology; D013116:Spinal Cord / Q000166:cytology* / Q000502:physiology; D013997:Time Factors
D003470:Culture Media; D007328:Insulin; D011700:Putrescine