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The group focuses on using microfabrication techniques to create novel high-throughput platforms that enable highly controlled environments for single neural cell studies. Our work is mainly focused on axonal regeneration, glial co-culture studies, and neuronal stem cell biology.
Prior efforts to study axons of the central and peripheral nervous systems in vitro have been limited by poor viability of low-density neuronal cultures and an inability to compartmentalize axons from neuronal cell bodies. To this extent, our group has developed a suite of microfluidic platforms to allow for high-throughput analysis of axon response to a variety of degenerative/protective cues (Fig. 1), selective myelination through axon-glia co-culture (Fig. 2), and compartmentalization of extending processes from organotypic tissue explants, among many other neuroscience applications. Using these platforms, we can quantitatively assess several parameters of axon degeneration, including morphology and transport in individual axons.
Fig. 1: Multiple experiments can be performed in the circular platform. Four consecutive labeling experiments (i) Control, (ii) 5uM CalceinAM (Green), (iii) 5 uM Cell Tracker Red, (iv) 5 uM CalceinAM (Green) were performed in a fully segmented device configuration (top right). The top image shows the entire experimental area and each inset (i-iv) shows magnified views of each experiment. Dye introduced into the axonal compartments entered axons and retrogradely labeled neuronal cell bodies.
Fig. 2: (A) The axons extended through the microchannels between the two compartments into the axon/glia compartment. Calcein Green staining. (B-E) are images of axon/glia compartment at two weeks in co-culture demonstrate that the axons grew from the soma compartment into the axon/glia compartment through the axon-guiding microchannels but dendrites and neuronal soma could not reach into the axon/glia compartment. Double immunoflurescence staining of axons (anti-neurofilament, NF, red and myelinated OLs (anti-MBP, green).
In Hong Yang, PhD
Suneil Hosmane, MS
Rezina Siddique, MS
Arun Venkatesan, MD, PhD - Johns Hopkins School of Medicine (Neurology)
Thomas Brushart, MD - Johns Hopkins School of Medicine (Orthopedic Surgery)
John Mcdonald, MD, PhD - Kennedy Krieger Institute
Ahmet Hoke, MD, PhD - Johns Hopkins School of Medicine (Neurology)
Avindra Nath, MD - Johns Hopkins School of Medicine (Neurology)
Fengquan Zhou, PhD - Johns Hopkins School of Medicine (Orthopedic Surgery)
JHU Institute of NanoBioTechnology 1207340082, Junior Facutly Pilot Award, Bio Medicine Pilot Award
NIH National Research Council Award (NRSA) F31NS066753-02
Maryland Stem Cell Fund
Department of Defense BC102629, MS100367
Ziegler L, Grigoryan S, Yang IH, Thakor N, Goldstein RS, Efficient Generation of Schwann Cells from Human Embryonic Stem Cell-Derived Neurospheres, Stem Cell Review Report, in press

Hosmane S, Yang I, Ruffin A, Thakor N, Venkatesan A, Circular compartmentalized microfluidic platform: Study of axon-glia interactions, Lab Chip, 10:741-7, 2010 [text]

Yang IH, Siddique R, Hosmane S, Thakor NV, and Hoke A, Compartmentalized microfluidic culture platform to study mechanism, Exp Neurol, 218(1):124-8, 2009 [text]

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