Substrate Micropatterning as a New in Vitro Cell Culture System to Study Myelination

Abstract

Myelination is a highly regulated developmental process whereby oligodendrocytes in the central nervous system and Schwann cells in the peripheral nervous system ensheathe axons with a multilayered concentric membrane. Axonal myelination increases the velocity of nerve impulse propagation. In this work, we present a novel in vitro system for coculturing primary dorsal root ganglia neurons along with myelinating cells on a highly restrictive and micropatterned substrate. In this new coculture system, neurons survive for several weeks, extending long axons on defined Matrigel tracks. On these axons, myelinating cells can achieve robust myelination, as demonstrated by the distribution of compact myelin and nodal markers. Under these conditions, neurites and associated myelinating cells are easily accessible for studies on the mechanisms of myelin formation and on the effects of axonal damage on the myelin sheath.

Myelination and node of Ranvier formation on sensory neurons in a defined in vitro system

“…Myelination of neurons by Schwann cells has been extensively studied using dorsal root ganglia (DRG) cultures in a variety of serum containing and serum-free in vitro systems, with most systems relying on the use of biological substrates for cell culture (Callizot et al, 2011; Hlady and Hodgkinson, 2007; Liazoghli et al, 2011; Svenningsen et al, 2003; Wood et al, 1990). Less common has been node of Ranvier formation by Schwann cells on motoneurons (Pang et al, 2012; Rumsey et al, 2009) and by oligodendrocytes on CNS neurons (Pang et al, 2012) in in vitro systems.…”

Microfluidic probes for use in life sciences and medicine

“…8a; this method could be used to study and monitor myelination in vitro. 37 Similarly, a single axon of a live hippocampal neuron was perfused with 5 Biopatterning with the MFP. (a) Fluorescence micrograph of an array of two different proteins patterned using the MFP.…”

Highly reproducible quantification of apoptotic cells using micropatterned culture of neurons

“…The micropatterning of cells can be achieved in numerous ways such as photolithography, soft lithography, and the stencil technique [7][8][9][10] technique relies on the use of a stencil with well-defined patterns for the localization of cell culture and adhesion. A stencil is a thin membrane consisting of perforated patterns.…”