title: A new long term in vitro model of myelination.
abstract: Besides in vivo models, co-cultures systems making use of Rat dorsal root ganglion explants/Schwann cells (SC) are widely used to essentially study myelination in vitro. In the case of animal models of demyelinating diseases, it is expected to reproduce a pathological process; conversely the co-cultures are primarily developed to study the myelination process and in the aim to use them to replace animals in experiences of myelin destruction or functional disturbances. We describe (in terms of protein expression kinetic) a new in vitro model of sensory neurons/SC co-cultures presenting the following advantages: both sensory neurons and SC originate from the same individual; sensory neurons and SC being dissociated, they can be co-cultured in monolayer, allowing an easier microscope observation; the co-culture can be maintained in a serum-free medium for at less three months, allowing kinetic studies of myelin formation both at a molecular and cellular level. Optimizing culture conditions permits to use 96-well culture plates; image analyses conducted with an automatic image analyzer allows rapid, accurate and quantitative expression of results. Finally, this system was proved by measuring the apparition of myelin protein to mimic in vitro the physiological process of in vivo myelination.
authors.affiliation: Neuron Experts SAS, Faculté de Médecine Nord, 51 Boulevard Pierre Dramard, Marseille Cedex 20, France. noelle.callizot@neuronexperts.com
authors.forename: Noelle
authors.initials: N
authors.lastname: Callizot
authors.forename: Maud
authors.initials: M
authors.lastname: Combes
authors.forename: Rémy
authors.initials: R
authors.lastname: Steinschneider
authors.forename: Philippe
authors.initials: P
authors.lastname: Poindron
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques / Q000379:methods; D017630:Connexins / Q000378:metabolism; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D005727:Ganglia, Spinal / Q000166:cytology; D015870:Gene Expression / Q000502:physiology; D007700:Kinetics; D004676:Myelin Basic Protein / Q000378:metabolism; D018993:Myelin P0 Protein / Q000378:metabolism; D009185:Myelin Proteins / Q000378:metabolism; D009186:Myelin Sheath / Q000502:physiology*; D018992:Myelin-Associated Glycoprotein / Q000378:metabolism; D020932:Nerve Growth Factor / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012583:Schwann Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D011984:Sensory Receptor Cells / Q000166:cytology*; D044967:Serum / Q000502:physiology; D000097002:Gap Junction beta-1 Protein
chemical_list: D001205:Ascorbic Acid; D015816:Cell Adhesion Molecules, Neuronal; D017630:Connexins; D016895:Culture Media, Serum-Free; D004676:Myelin Basic Protein; D018993:Myelin P0 Protein; D009185:Myelin Proteins; D018992:Myelin-Associated Glycoprotein; D020932:Nerve Growth Factor; D000097002:Gap Junction beta-1 Protein; C105177:Cntnap1 protein, rat; C436006:Mbp protein, rat; C560548:Mpz protein, rat; C489546:Pmp22 protein, rat