Cytological, molecular, cytogenetic, and physiological characterization of a novel immortalized human enteric glial cell line
Enteric glial cells (EGCs), the major components of the enteric nervous system (ENS), are implicated in the maintenance of gut homeostasis, thereby leading to severe pathological conditions when impaired. However, due to technical difficulties associated with EGCs isolation and cell culture maintena...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-04-01
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| Series: | Frontiers in Cellular Neuroscience |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fncel.2023.1170309/full |
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| author | Lisa Zanoletti Lisa Zanoletti Aurora Valdata Kristina Nehlsen Pawan Faris Pawan Faris Claudio Casali Rosalia Cacciatore Ilaria Sbarsi Francesca Carriero Davide Arfini Lies van Baarle Veronica De Simone Giulia Barbieri Elena Raimondi Tobias May Francesco Moccia Mauro Bozzola Gianluca Matteoli Sergio Comincini Federico Manai |
| author_facet | Lisa Zanoletti Lisa Zanoletti Aurora Valdata Kristina Nehlsen Pawan Faris Pawan Faris Claudio Casali Rosalia Cacciatore Ilaria Sbarsi Francesca Carriero Davide Arfini Lies van Baarle Veronica De Simone Giulia Barbieri Elena Raimondi Tobias May Francesco Moccia Mauro Bozzola Gianluca Matteoli Sergio Comincini Federico Manai |
| author_sort | Lisa Zanoletti |
| collection | DOAJ |
| description | Enteric glial cells (EGCs), the major components of the enteric nervous system (ENS), are implicated in the maintenance of gut homeostasis, thereby leading to severe pathological conditions when impaired. However, due to technical difficulties associated with EGCs isolation and cell culture maintenance that results in a lack of valuable in vitro models, their roles in physiological and pathological contexts have been poorly investigated so far. To this aim, we developed for the first time, a human immortalized EGC line (referred as ClK clone) through a validated lentiviral transgene protocol. As a result, ClK phenotypic glial features were confirmed by morphological and molecular evaluations, also providing the consensus karyotype and finely mapping the chromosomal rearrangements as well as HLA-related genotypes. Lastly, we investigated the ATP- and acetylcholine, serotonin and glutamate neurotransmitters mediated intracellular Ca2+ signaling activation and the response of EGCs markers (GFAP, SOX10, S100β, PLP1, and CCL2) upon inflammatory stimuli, further confirming the glial nature of the analyzed cells. Overall, this contribution provided a novel potential in vitro tool to finely characterize the EGCs behavior under physiological and pathological conditions in humans. |
| first_indexed | 2024-04-09T17:06:13Z |
| format | Article |
| id | doaj.art-46085daa87684a25bcbdff56101104a8 |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-04-09T17:06:13Z |
| publishDate | 2023-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-46085daa87684a25bcbdff56101104a82023-04-20T13:25:00ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022023-04-011710.3389/fncel.2023.11703091170309Cytological, molecular, cytogenetic, and physiological characterization of a novel immortalized human enteric glial cell lineLisa Zanoletti0Lisa Zanoletti1Aurora Valdata2Kristina Nehlsen3Pawan Faris4Pawan Faris5Claudio Casali6Rosalia Cacciatore7Ilaria Sbarsi8Francesca Carriero9Davide Arfini10Lies van Baarle11Veronica De Simone12Giulia Barbieri13Elena Raimondi14Tobias May15Francesco Moccia16Mauro Bozzola17Gianluca Matteoli18Sergio Comincini19Federico Manai20Department of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyDepartment of Chronic Diseases and Metabolism (CHROMETA), KU Leuven, Leuven, BelgiumDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyInSCREENeX GmbH, Braunschweig, GermanyDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyDepartment of Biology, College of Science, Salahaddin University-Erbil, Erbil, IraqDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyImmunohematology and Transfusion Service, I.R.C.C.S. Policlinico San Matteo, Pavia, ItalyImmunohematology and Transfusion Service, I.R.C.C.S. Policlinico San Matteo, Pavia, ItalyDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyDepartment of Chronic Diseases and Metabolism (CHROMETA), KU Leuven, Leuven, BelgiumDepartment of Chronic Diseases and Metabolism (CHROMETA), KU Leuven, Leuven, BelgiumDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyInSCREENeX GmbH, Braunschweig, GermanyDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyUniversity of Pavia, Pavia, ItalyDepartment of Chronic Diseases and Metabolism (CHROMETA), KU Leuven, Leuven, BelgiumDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyDepartment of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Pavia, ItalyEnteric glial cells (EGCs), the major components of the enteric nervous system (ENS), are implicated in the maintenance of gut homeostasis, thereby leading to severe pathological conditions when impaired. However, due to technical difficulties associated with EGCs isolation and cell culture maintenance that results in a lack of valuable in vitro models, their roles in physiological and pathological contexts have been poorly investigated so far. To this aim, we developed for the first time, a human immortalized EGC line (referred as ClK clone) through a validated lentiviral transgene protocol. As a result, ClK phenotypic glial features were confirmed by morphological and molecular evaluations, also providing the consensus karyotype and finely mapping the chromosomal rearrangements as well as HLA-related genotypes. Lastly, we investigated the ATP- and acetylcholine, serotonin and glutamate neurotransmitters mediated intracellular Ca2+ signaling activation and the response of EGCs markers (GFAP, SOX10, S100β, PLP1, and CCL2) upon inflammatory stimuli, further confirming the glial nature of the analyzed cells. Overall, this contribution provided a novel potential in vitro tool to finely characterize the EGCs behavior under physiological and pathological conditions in humans.https://www.frontiersin.org/articles/10.3389/fncel.2023.1170309/fullenteric glial cellsenteric nervous systemtransgene immortalizationviral transductionimmortalized human cell line |
| spellingShingle | Lisa Zanoletti Lisa Zanoletti Aurora Valdata Kristina Nehlsen Pawan Faris Pawan Faris Claudio Casali Rosalia Cacciatore Ilaria Sbarsi Francesca Carriero Davide Arfini Lies van Baarle Veronica De Simone Giulia Barbieri Elena Raimondi Tobias May Francesco Moccia Mauro Bozzola Gianluca Matteoli Sergio Comincini Federico Manai Cytological, molecular, cytogenetic, and physiological characterization of a novel immortalized human enteric glial cell line Frontiers in Cellular Neuroscience enteric glial cells enteric nervous system transgene immortalization viral transduction immortalized human cell line |
| title | Cytological, molecular, cytogenetic, and physiological characterization of a novel immortalized human enteric glial cell line |
| title_full | Cytological, molecular, cytogenetic, and physiological characterization of a novel immortalized human enteric glial cell line |
| title_fullStr | Cytological, molecular, cytogenetic, and physiological characterization of a novel immortalized human enteric glial cell line |
| title_full_unstemmed | Cytological, molecular, cytogenetic, and physiological characterization of a novel immortalized human enteric glial cell line |
| title_short | Cytological, molecular, cytogenetic, and physiological characterization of a novel immortalized human enteric glial cell line |
| title_sort | cytological molecular cytogenetic and physiological characterization of a novel immortalized human enteric glial cell line |
| topic | enteric glial cells enteric nervous system transgene immortalization viral transduction immortalized human cell line |
| url | https://www.frontiersin.org/articles/10.3389/fncel.2023.1170309/full |
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