Neuronal Network Dynamics in 2D and 3D in vitro Neuroengineered Systems

151 Pages·2016·8.146 MB·English

by  Monica Frega (auth.)

Springer Theses
Recognizing Outstanding Ph.D. Research
Monica Frega

Neuronal Network
Dynamics in
2D and 3D in vitro
Neuroengineered
Systems
Springer Theses
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Monica Frega
Neuronal Network Dynamics
in 2D and 3D in vitro
Neuroengineered Systems
Doctoral Thesis accepted by
the University of Genoa, Italy
123
Author Supervisor
Monica Frega Prof. SergioMartinoia
Department ofInformatics, Bioengineering, Department ofInformatics, Bioengineering,
Robotics andSystem Engineering Robotics andSystem Engineering
University of Genoa University of Genoa
Genoa Genoa
Italy Italy
ISSN 2190-5053 ISSN 2190-5061 (electronic)
SpringerTheses
ISBN978-3-319-30236-2 ISBN978-3-319-30237-9 (eBook)
DOI 10.1007/978-3-319-30237-9
LibraryofCongressControlNumber:2016932514
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Part of this thesis have been published in the following articles:
Peer-reviewed International Journals
[J.1] Tedesco, M., Frega, M., Martinoia, s., Pesce, M., Massobrio, P. (2015).
Interfacing 3D engineered neuronal cultures to Micro-Electrode Arrays: an inno-
vative in vitro experimental model. Journal of Visualized Experiments (104),
e53080, doi: 10.3791/53080.
[J.2] Frega, M., Tedesco, M., Massobrio, P., Pesce, M., Martinoia, S. (2014).
Enhanced network dynamics in 3D engineered neuronal cultures: a new experi-
mental model for network electrophysiology. Scientific Reports, 4, 5489.
[J.3] Colombi,I.,Mahajani,S.,Frega,M.,Gasparini,L.,Chiappalone,M.(2013).
Effects of antiepileptic drugs on hippocampal neurons coupled to micro-electrode
arrays. Frontiers in Neuroengineering, 6, 10.
[J.4] Frega,M.,Pasquale,V.,Tedesco,M.,Marcoli,M.,Contestabile,A.,Nanni,
M., Bonzano, L., Maura, G., Chiappalone, M. (2012). Cortical cultures coupled to
Micro-Electrode Arrays: A novel approach to perform in vitro excitotoxicity test-
ing. Neurotoxicology and teratology, 34(1), 116-127.
Peer-reviewed conference proceedings
[C.1] Tedesco, M., Frega, M., Pastorino, L. Massobrio, M., Martinoia, S. 3D
engineered neural networks coupled to Micro-Electrode based devices: a new
experimental model for neurophysiological applications. AISEM Annual
Conference, 2015 XVIII, 3rd–5th February 2015, Trento, Italy.
[C.2] Frega, M., Tedesco, M., Massobrio, P., Pesce, M., Martinoia, S. 3Dneural
networks coupled to planar microtransducer arrays: an innovative in-vitro experi-
mental model for investigating network dynamics. 9th International Meeting on
Substrate-Integrated Mircoelectrode Arrays, 1st–4th July 2014, Reutlingen,
Germany.
[C.3] Frega, M., Tedesco, M., Massobrio, P., Martinoia, S. 3D neural networks
coupledtoMicroElectrodeArrays:anewexperimentalmodelforneuro-electronic
interfaces.4thcongress-NationalGroupofBioengineering(GNB),25th–27thJune
2014, Pavia (Italy).
[C.4] Frega,M.,Tedesco,M.,Massobrio,P.,Pesce,M.,Williamson,A.,Schober,
A., Martinoia, S. 3D Engineered Neural Networks Coupled to Micro-Electrode
Arrays: Development of an Innovative In-Vitro Experimental Model for
Neurophysiological Studies. 6th International IEEE/EMBS Conference on Neural
Engineering (NER), 5th–8th November 2013, San Diego (USA).
[C.5] Frega, M., Tedesco, M., Massobrio, P., Pesce, M., Martinoia, S. 3D neu-
ronal networks coupled to Micro-Electrode Arrays: an innovative in vitro experi-
mental model to study network dynamics. 8th International Meeting on
Substrate-Integrated Microelectrode Arrays (SIMEA 2010), 10th–13th July 2012,
Reutlingen (Germany).
Abstract to National and International conferences
[A.1] Tedesco, M., Frega, M., Massobrio, P., Pesce, M., Martinoia, S. 3D neu-
ronal cultures coupled to multi-electrode arrays: a new experimental model
forin-vitroelectrophysiology.9thForumofEuropeanNeuroscience(FENS2014),
5th–9th July 2014, Milan (Italy).
[A.2] Frega,M., Tedesco, M., Massobrio, P., Martinoia,S. Comparison between
the electrophysiological activity of 2D and 3D hippocampal neural networks cou-
pledtomicro-electrodearrays.9thForumofEuropeanNeuroscience(FENS2014),
5th–9th July 2014, Milan (Italy).
[A.3] Frega, M., Tedesco, M., Massobrio, P., Pesce, M., Martinoia, S.
Comparison between electrophysiological activity of 2D and 3D cultured hip-
pocampal networks coupled to micro-electrode arrays. Neuroscience, 9th–13th
November 2013, San Diego (USA).
[A.4] Martinoia, S., Tedesco, M., Frega, M., Massobrio, P., Wiliamson, A.,
Schober, A. 3-D neuronal cultures coupled to 3-D multi-electrode arrays: a new
biohybrid investigational platform for network neurophysiology. Neuroscience,
9th–13th November 2013, San Diego (USA).
[A.5] Martinoia, S., Frega, M., Tedesco, M., Pesce, M., Massobrio, P. 3D neu-
ronal networks coupled to Micro-Electrode Arrays: development of an in vitro
experimental model to study network dynamics. 8th Forum of European
Neuroscience (FENS 2012), 14th–18th July 2012, Barcelona (Spain).
’
Supervisor s Foreword
It is a great pleasure to introduce Dr. Monica Frega’s thesis work accepted for
publication within Springer Theses and awarded with a prize for outstanding
originalwork.MonicaFregaperformedherMasterthesisundermysupervisionand
then joined my research group on Neuroengineering and Bionanotechnologies
(Department of Informatics, Bioengineering, Robotics and Systems Engineering)
for her Ph.D. program on Bioengineering at the University of Genova. She started
in January 2011 and completed her studies with a successful thesis defense on
April 4, 2014.
The thesis reports an innovative study in the design and implementation of
new-engineered three-dimensional (3D) neuronal cultures coupled to micro elec-
trode arrays (MEAs). Part of the presented results have been awarded by the
NationalBioengineeringGroupwiththe“Awardforyoungscientists”in2014and
most of them have been published in Scientific Report (Nature Publishing Group)
on June 30, 2014.
The use of in vitro neuronal networks for neurophysiological investigation is a
well-established technique since decades. It is widely used for molecular, cellular,
and networks studies, mainly for its possibility of manipulation, observation,
control, and long-term culturing. The coupling of such in vitro models, derived
from primary cultures of rodents (embryo or neonatal), to MEAs technology goes
back to the’70s. These model networks are routinely grown onto planar substrate;
attempts in improving this approach are concentrated on developing new tech-
nologies for recording and stimulating the electrophysiological activity of such
experimental preparations. Inthe past decade, there was increasing attention tothe
development of 3D cultured networks simply by recognizing the advantages, from
the physiological point of view, of such in vitro preparations capable of better
mimicking and recapitulating the in vivo situation. Notwithstanding the increased
momentumin3Dnetworks,alltheelectrophysiologicalstudiesatthenetworklevel
arestillbasedontwo-dimensional(2D)invitromodels(i.e.,networksgrownontoa
planar substrate).
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viii Supervisor’sForeword
In her thesis work, Monica Frega greatly contributed to the development of a
new experimental paradigm constituted by 3D neural networks coupled to micro
electrode arrays (MEAs). These 3D networks were systematically characterized
fromanexperimentalpointofview,andthenetworkdynamicsdeeplyinvestigated,
analyzed, and compared with standard 2D networks. Her work included experi-
mental design, data collection, and specific data processing. For the first time,
Monica Frega demonstrated the possibility of coupling 3D cultured networks to
planar MEAs, and verified the capability of such 3D networks of showing a dif-
ferentelectrophysiologicalactivity(withrespectto2Dsister-cultures)withspecific
features and with much more similarities to the in vivo situation.
This new approach opens new avenues in the network electrophysiology field,
allowing studying in vitro neuronal systems in a more physiological condition,
approaching and mimicking natural in vivo assemblies. Moreover, it constitutes a
step forward in the field of neuroengineering, with specific relevance for the
development of new cellular, tissue-like in vitro experimental model systems with
future far-reaching implications in the field of in vivo neural interface and neural
grafting.
Genoa Prof. Sergio Martinoia
November 2015
Acknowledgments
DuringmyPh.D.Ihadtheoccasiontogodeeperintomyinterestforresearchandat
theendoftheseyearsmypassiongraduallygrew.Thispersonalgrowthwouldnot
have been possible without competent and professional people who were really
passionateintheirworkandhumanlyinteresting.Iwouldliketothankthesepeople
one by one. First of all, I express my sincere gratitude to my supervisor Sergio
Martinoia. This work could not be performed without his valuable guidance and
trust.IalsothankPaoloMassobrio.Workingwiththetwoofyouwasgreat.Ireally
enjoyed my research, and you strongly supported and encouraged my enthusiasm.
I was sharing with both of you not only scientific ideas and results, but also my
personallife.Youwerenotonlymysupervisors,butgreatfriendsofmine.Without
such a good relationship, I am sure that the outcome of my Ph.D. would not have
been the same, both in terms of “results” and in terms of my personal growth.
I thank MariaTeresa Tedesco: you provided me a lot of support and help and you
conveyed tomethepassionfor yourwork.Ialsothank MichelaChiappalone:you
alwayssupportedmeandyouwereimportantformyscientificandpersonalgrowth.
IthankVirginia,Andrea,andDaniele:withyouIsharedmostofthetimeduring
my Ph.D. We did not share only scientific ideas, but also many nice moments and
interesting talks together.
ThesethreeyearsofPh.D.werealongjourney:Icouldnothavecompletedthis
journey without the support of my friends and family. I thank all of you!
In particular, I thank the friends with whom I have shared almost everything:
Albertone,Giulia,Davide,Andrea,Luca,Ignazio,Chiusky,Marialaura,Anna,and
Susanna. You were great and helpful company: I shared my life and the important
steps of my personal development with you. It is incredible how our friendship
grew with time. I have no words to express my gratitude.
Andofcourse,abigthankyoutoMattia.Youalwaysprovidemesupport,help,
and encouragement. You are a gift to me: you really changed my life in this year.
Iamimpressedbyhowwewereabletofacedifficultiesandhowthedistancemade
uscloserandmadeourrelationshipstronger.Iamreallygratefultohaveyouatmy
side. And the best is yet to come!
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