{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/90854"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/90854","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Characterization of spontaneous and evoked activity of mouse embryonic stem cell derived motor neurons using optogenetics and micro electrode array electrophysiology","abstract":"Thorough characterization of distinct neuronal lineages derived from progenitor cells is essential for the development of biological models that can recreate native function of neurons of the central nervous system while permitting researchers to have an easily accessible source of cellular building blocks. These models are particularly important for studies on neurological disorders aimed at understanding disease pathways and for high throughput testing of drug candidates. Furthermore, developing a model system with stem cell-derived spinal cord motor neurons (MN) is attractive for designing control systems for soft-tissue robotics. The critical first step towards building a useful model with MNs require a detail investigation of spontaneous and stimulation-evoked electrical activity of developing MN networks. The following work uses multi-electrode array (MEA) electrophysiology and optogenetics to characterize electrical activity of MNs differentiated from mouse embryonic stem (mES) cells. A customizable substrate integrated 60-electrode MEA chip was designed for this study. MEA chips were fabricated with platinum deposited on borofloat glass for detection of small changes in electrical field potentials resulting from neuronal activity that causes small change in ionic currents. A Multi-Channel Systems amplifier was used for recording. MN embryoid bodies (MEBs) were grown in these MEA chips, and differentiation of mES cells into MNs was monitored by the expression of eGFP with a MN specific promoter, Hb9. The mES cell line was transfected with channelrhodopsin-2 (ChR2) tagged with Td-Tomato which allowed for optogenetic stimulation of the networks with a 470 nm LED. Network firing patterns were evaluated for bursting activity and spectral content using analysis algorithms developed in MATLAB. Data presented here demonstrated that MEBs are spontaneously active, they develop a robust network synchronization, and optogenetic stimulation increased the firing rate and affected the firing patterns. This work established a model system with mES cell derived MNs. These findings are a milestone in the efforts of developing neural circuits that can be used to potentially control higher order soft-tissue robotics.","abstract_html":"Thorough characterization of distinct neuronal lineages derived from progenitor cells is essential for the development of biological models that can recreate native function of neurons of the central nervous system while permitting researchers to have an easily accessible source of cellular building blocks. These models are particularly important for studies on neurological disorders aimed at understanding disease pathways and for high throughput testing of drug candidates. Furthermore, developing a model system with stem cell-derived spinal cord motor neurons (MN) is attractive for designing control systems for soft-tissue robotics. The critical first step towards building a useful model with MNs require a detail investigation of spontaneous and stimulation-evoked electrical activity of developing MN networks. The following work uses multi-electrode array (MEA) electrophysiology and optogenetics to characterize electrical activity of MNs differentiated from mouse embryonic stem (mES) cells. A customizable substrate integrated 60-electrode MEA chip was designed for this study. MEA chips were fabricated with platinum deposited on borofloat glass for detection of small changes in electrical field potentials resulting from neuronal activity that causes small change in ionic currents. A Multi-Channel Systems amplifier was used for recording. MN embryoid bodies (MEBs) were grown in these MEA chips, and differentiation of mES cells into MNs was monitored by the expression of eGFP with a MN specific promoter, Hb9. The mES cell line was transfected with channelrhodopsin-2 (ChR2) tagged with Td-Tomato which allowed for optogenetic stimulation of the networks with a 470 nm LED. Network firing patterns were evaluated for bursting activity and spectral content using analysis algorithms developed in MATLAB. Data presented here demonstrated that MEBs are spontaneously active, they develop a robust network synchronization, and optogenetic stimulation increased the firing rate and affected the firing patterns. This work established a model system with mES cell derived MNs. These findings are a milestone in the efforts of developing neural circuits that can be used to potentially control higher order soft-tissue robotics.","abstract_has_math":false,"creators":["Pagán-Díaz, Gelson J"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"M.S.","degree_level":"Thesis","degree_discipline":"Bioengineering","degree_department":null,"school":null,"contributors":["Sengupta, Parijat","Bahisr, Rashid"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2016,"date_issued":"2016-07-07T20:35:31Z","date_published":"2016-07-07T20:35:31Z","updated_at":"2026-07-22T22:26:34Z","subjects":["Multi-electrode array (MEA)","microelectrode array","mES cells, embryoid bodies","electrophysiology"],"languages":["en"],"rights":["Copyright 2016 Gelson J. Pagan"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/90854","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Sengupta, Parijat","Bahisr, Rashid"]},{"key":"dc:creator","label":"Author","values":["Pagán-Díaz, Gelson J"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2016-07-07T20:35:31Z","2018-07-08T09:15:36Z","2016-04-29","2016-05"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Bioengineering"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M.S."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Multi-electrode array (MEA)","microelectrode array","mES cells, embryoid bodies","electrophysiology"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2016 Gelson J. Pagan"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/90854"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Thorough characterization of distinct neuronal lineages derived from progenitor cells is essential for the development of biological models that can recreate native function of neurons of the central nervous system while permitting researchers to have an easily accessible source of cellular building blocks. These models are particularly important for studies on neurological disorders aimed at understanding disease pathways and for high throughput testing of drug candidates. Furthermore, developing a model system with stem cell-derived spinal cord motor neurons (MN) is attractive for designing control systems for soft-tissue robotics. The critical first step towards building a useful model with MNs require a detail investigation of spontaneous and stimulation-evoked electrical activity of developing MN networks. The following work uses multi-electrode array (MEA) electrophysiology and optogenetics to characterize electrical activity of MNs differentiated from mouse embryonic stem (mES) cells. A customizable substrate integrated 60-electrode MEA chip was designed for this study. MEA chips were fabricated with platinum deposited on borofloat glass for detection of small changes in electrical field potentials resulting from neuronal activity that causes small change in ionic currents. A Multi-Channel Systems amplifier was used for recording. MN embryoid bodies (MEBs) were grown in these MEA chips, and differentiation of mES cells into MNs was monitored by the expression of eGFP with a MN specific promoter, Hb9. The mES cell line was transfected with channelrhodopsin-2 (ChR2) tagged with Td-Tomato which allowed for optogenetic stimulation of the networks with a 470 nm LED. Network firing patterns were evaluated for bursting activity and spectral content using analysis algorithms developed in MATLAB. Data presented here demonstrated that MEBs are spontaneously active, they develop a robust network synchronization, and optogenetic stimulation increased the firing rate and affected the firing patterns. This work established a model system with mES cell derived MNs. These findings are a milestone in the efforts of developing neural circuits that can be used to potentially control higher order soft-tissue robotics.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2018-05-01","The student, Gelson Pagán-Díaz, accepted the attached license on 2016-04-29 at 15:27.","The student, Gelson Pagán-Díaz, submitted this Thesis for approval on 2016-04-29 at 15:32.","This Thesis was approved for publication on 2016-04-29 at 16:53.","DSpace SAF Submission Ingestion Package generated from Vireo submission #9591 on 2016-07-07 at 13:51:15","Made available in DSpace on 2016-07-07T20:35:31Z (GMT). No. of bitstreams: 2 PAGAN-DIAZ-THESIS-2016.pdf: 7157479 bytes, checksum: 9a9660f2f6dcb05b1c52084bb2369f0b (MD5) LICENSE.txt: 4213 bytes, checksum: f8fe38dc326310a0b6b1a979302fc252 (MD5) Previous issue date: 2016-04-29","Embargo set by: Seth Robbins for item 93207 Lift date: 2018-07-07T20:35:34Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","U of I Only Restriction Lifted for Item 93207 on 2018-07-08T09:15:36Z."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Characterization of spontaneous and evoked activity of mouse embryonic stem cell derived motor neurons using optogenetics and micro electrode array electrophysiology"]}]}],"canonical_facts":{"dc:contributor":["Sengupta, Parijat","Bahisr, Rashid"],"dc:creator":["Pagán-Díaz, Gelson J"],"dc:date":["2016-07-07T20:35:31Z","2018-07-08T09:15:36Z","2016-04-29","2016-05"],"dc:description":["Thorough characterization of distinct neuronal lineages derived from progenitor cells is essential for the development of biological models that can recreate native function of neurons of the central nervous system while permitting researchers to have an easily accessible source of cellular building blocks. These models are particularly important for studies on neurological disorders aimed at understanding disease pathways and for high throughput testing of drug candidates. Furthermore, developing a model system with stem cell-derived spinal cord motor neurons (MN) is attractive for designing control systems for soft-tissue robotics. The critical first step towards building a useful model with MNs require a detail investigation of spontaneous and stimulation-evoked electrical activity of developing MN networks. The following work uses multi-electrode array (MEA) electrophysiology and optogenetics to characterize electrical activity of MNs differentiated from mouse embryonic stem (mES) cells. A customizable substrate integrated 60-electrode MEA chip was designed for this study. MEA chips were fabricated with platinum deposited on borofloat glass for detection of small changes in electrical field potentials resulting from neuronal activity that causes small change in ionic currents. A Multi-Channel Systems amplifier was used for recording. MN embryoid bodies (MEBs) were grown in these MEA chips, and differentiation of mES cells into MNs was monitored by the expression of eGFP with a MN specific promoter, Hb9. The mES cell line was transfected with channelrhodopsin-2 (ChR2) tagged with Td-Tomato which allowed for optogenetic stimulation of the networks with a 470 nm LED. Network firing patterns were evaluated for bursting activity and spectral content using analysis algorithms developed in MATLAB. Data presented here demonstrated that MEBs are spontaneously active, they develop a robust network synchronization, and optogenetic stimulation increased the firing rate and affected the firing patterns. This work established a model system with mES cell derived MNs. These findings are a milestone in the efforts of developing neural circuits that can be used to potentially control higher order soft-tissue robotics.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2018-05-01","The student, Gelson Pagán-Díaz, accepted the attached license on 2016-04-29 at 15:27.","The student, Gelson Pagán-Díaz, submitted this Thesis for approval on 2016-04-29 at 15:32.","This Thesis was approved for publication on 2016-04-29 at 16:53.","DSpace SAF Submission Ingestion Package generated from Vireo submission #9591 on 2016-07-07 at 13:51:15","Made available in DSpace on 2016-07-07T20:35:31Z (GMT). No. of bitstreams: 2 PAGAN-DIAZ-THESIS-2016.pdf: 7157479 bytes, checksum: 9a9660f2f6dcb05b1c52084bb2369f0b (MD5) LICENSE.txt: 4213 bytes, checksum: f8fe38dc326310a0b6b1a979302fc252 (MD5) Previous issue date: 2016-04-29","Embargo set by: Seth Robbins for item 93207 Lift date: 2018-07-07T20:35:34Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","U of I Only Restriction Lifted for Item 93207 on 2018-07-08T09:15:36Z."],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/2142/90854"],"dc:language":["en"],"dc:rights":["Copyright 2016 Gelson J. Pagan"],"dc:subject":["Multi-electrode array (MEA)","microelectrode array","mES cells, embryoid bodies","electrophysiology"],"dc:title":["Characterization of spontaneous and evoked activity of mouse embryonic stem cell derived motor neurons using optogenetics and micro electrode array electrophysiology"],"dc:type":["text"],"thesis:degree_discipline":["Bioengineering"],"thesis:degree_level":["Thesis"],"thesis:degree_name":["M.S."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:26:34Z"}