title: Generation of three-dimensional multiple spheroid model of olfactory ensheathing cells using floating liquid marbles.
abstract: We describe a novel protocol for three-dimensional culturing of olfactory ensheathing cells (OECs), which can be used to understand how OECs interact with other cells in three dimensions. Transplantation of OECs is being trialled for repair of the paralysed spinal cord, with promising but variable results and thus the therapy needs improving. To date, studies of OEC behaviour in a multicellular environment have been hampered by the lack of suitable three-dimensional cell culture models. Here, we exploit the floating liquid marble, a liquid droplet coated with hydrophobic powder and placed on a liquid bath. The presence of the liquid bath increases the humidity and minimises the effect of evaporation. Floating liquid marbles allow the OECs to freely associate and interact to produce OEC spheroids with uniform shapes and sizes. In contrast, a sessile liquid marble on a solid surface suffers from evaporation and the cells aggregate with irregular shapes. We used floating liquid marbles to co-culture OECs with Schwann cells and astrocytes which formed natural structures without the confines of gels or bounding layers. This protocol can be used to determine how OECs and other cell types associate and interact while forming complex cell structures.
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Brisbane, QLD 4111, Australia.
authors.forename: Raja K
authors.initials: RK
authors.lastname: Vadivelu
authors.affiliation: QLD Micro- and Nanotechnology Centre, Griffith University, 4111, Australia.
authors.forename: Chin H
authors.initials: CH
authors.lastname: Ooi
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Brisbane, QLD 4111, Australia.
authors.forename: Rebecca-Qing
authors.initials: RQ
authors.lastname: Yao
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Brisbane, QLD 4111, Australia.
authors.forename: Johana
authors.initials: J
authors.lastname: Tello Velasquez
authors.affiliation: Nature Communications, New York, USA.
authors.forename: Erika
authors.initials: E
authors.lastname: Pastrana
authors.affiliation: Centro de Biología Molecular Severo Ochoa &CIBERER, Madrid, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Diaz-Nido
authors.affiliation: Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: Filip
authors.initials: F
authors.lastname: Lim
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Brisbane, QLD 4111, Australia.|School of Biomedical Sciences, Queensland University of Technology, Brisbane, 4000, QLD, Australia.
authors.forename: Jenny A K
authors.initials: JA
authors.lastname: Ekberg
authors.affiliation: QLD Micro- and Nanotechnology Centre, Griffith University, 4111, Australia.
authors.forename: Nam-Trung
authors.initials: NT
authors.lastname: Nguyen
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Brisbane, QLD 4111, Australia.
authors.forename: James A
authors.initials: JA
authors.lastname: St John
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques / Q000379:methods*; D002450:Cell Communication / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D051379:Mice; D044085:Microfluidics / Q000379:methods; D009457:Neuroglia / Q000166:cytology* / Q000502:physiology; D009830:Olfactory Bulb / Q000166:cytology* / Q000502:physiology; D066330:Printing, Three-Dimensional; D018874:Spheroids, Cellular / Q000166:cytology* / Q000502:physiology