Myelination in mouse dorsal root ganglion/Schwann cell cocultures.
The established protocols for in vitro studies of peripheral nerve myelination with rat embryonic dorsal root ganglia (DRG) and postnatal Schwann cell cocultures do not work with mouse cells. Consequently, the full potential of this model, which would allow to perform cell type-specific, mixed genotype cocultures without cross-breeding the animals, cannot be exploited. We determined the conditions required to promote full myelination in cocultures of pre-purified mouse embryonic DRG and neonatal Schwann cells, and present a method which consistently yields 50-200 mature myelin sheaths/culture. Causes for the failure of the existing protocols to yield satisfactory results with mouse cells fell into three categories: the lack of adherent support provided by the substratum, growth factor and hormone deficiencies, and the high serum content of the media. For optimal results, mouse cocultures require a 3-dimensional substratum, a myelination-promoting culture medium containing pituitary extract, N2 supplement and forskolin, and a low serum concentration.
Päiväläinen
Satu
S
Department of Anatomy and Cell Biology, University of Oulu, Aapistie 7A, 90014 Oulu, Finland.
Nissinen
Marja
M
Honkanen
Henrika
H
Lahti
Outi
O
Kangas
Salla M
SM
Peltonen
Juha
J
Peltonen
Sirkku
S
Heape
Anthony M
AM
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D018920:Coculture Techniques; D004622:Embryo, Mammalian; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology*; D005786:Gene Expression Regulation / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D004676:Myelin Basic Protein / Q000378:metabolism*; D018992:Myelin-Associated Glycoprotein / Q000378:metabolism*; D009474:Neurons / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000502:physiology*; D012584:Sciatic Nerve / Q000166:cytology; D013997:Time Factors
D004676:Myelin Basic Protein; D018992:Myelin-Associated Glycoprotein