title: Axon myelination and electrical stimulation in a microfluidic, compartmentalized cell culture platform.
abstract: Axon demyelination contributes to the loss of sensory and motor function following injury or disease in the central nervous system. Numerous reports have demonstrated that myelination can be achieved in neuron/oligodendrocyte co-cultures. However, the ability to selectively treat neuron or oligodendrocyte (OL) cell bodies in co-cultures improves the value of these systems when designing mechanism-based therapeutics. We have developed a microfluidic-based compartmentalized culture system to achieve segregation of neuron and OL cell bodies while simultaneously allowing the formation of myelin sheaths. Our microfluidic platform allows for a high replicate number, minimal leakage, and high flexibility. Using a custom built lid, fit with platinum electrodes for electrical stimulation (10-Hz pulses at a constant 3 V with ~190 kΩ impedance), we employed the microfluidic platform to achieve activity-dependent myelin segment formation. Electrical stimulation of dorsal root ganglia resulted in a fivefold increase in the number of myelinated segments/mm² when compared to unstimulated controls (19.6 ± 3.0 vs. 3.6 ± 2.3 MBP+ segments/mm²). This work describes the modification of a microfluidic, multi-chamber system so that electrical stimulation can be used to achieve increased levels of myelination while maintaining control of the cell culture microenvironment.
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Johns Hopkins University, Baltimore, MD, USA.
authors.forename: In Hong
authors.initials: IH
authors.lastname: Yang
authors.forename: Devin
authors.initials: D
authors.lastname: Gary
authors.forename: Misti
authors.initials: M
authors.lastname: Malone
authors.forename: Stephen
authors.initials: S
authors.lastname: Dria
authors.forename: Thierry
authors.initials: T
authors.lastname: Houdayer
authors.forename: Visar
authors.initials: V
authors.lastname: Belegu
authors.forename: John W
authors.initials: JW
authors.lastname: McDonald
authors.forename: Nitish
authors.initials: N
authors.lastname: Thakor
mesh_terms: D000818:Animals; D001369:Axons / Q000378:metabolism* / Q000648:ultrastructure; D018929:Cell Culture Techniques / Q000295:instrumentation*; D018920:Coculture Techniques / Q000295:instrumentation; D004558:Electric Stimulation; D005727:Ganglia, Spinal / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009186:Myelin Sheath / Q000378:metabolism*; D009836:Oligodendroglia / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley