Primary Cortical Cell Tri-Culture-Based Screening of Neuroinflammatory Response in Toll-like Receptor Activation
The activation of toll-like receptors (TLRs) in the central nervous system (CNS) can lead to neuroinflammation and contribute to many neurological disorders, including autoimmune diseases. Cell culture models are powerful tools for studying specific molecular and cellular mechanisms that contribute...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-08-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/10/9/2122 |
| _version_ | 1797490974283268096 |
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| author | Noah Goshi Hyehyun Kim Erkin Seker |
| author_facet | Noah Goshi Hyehyun Kim Erkin Seker |
| author_sort | Noah Goshi |
| collection | DOAJ |
| description | The activation of toll-like receptors (TLRs) in the central nervous system (CNS) can lead to neuroinflammation and contribute to many neurological disorders, including autoimmune diseases. Cell culture models are powerful tools for studying specific molecular and cellular mechanisms that contribute to these disease states and identifying potential therapeutics. However, most cell culture models have limitations in capturing biologically relevant phenomena, due in part to the non-inclusion of necessary cell types. Neurons, astrocytes, and microglia (critical cell types that play a role in neuroinflammation) all express at least a subset of TLRs. However, the response of each of these cell types to various TLR activation, along with their relative contribution to neuroinflammatory processes, is far from clear. In this study, we demonstrate the screening capabilities of a primary cortical cell tri-culture of neuron, astrocyte, and microglia from neonatal rats. Specifically, we compare the neuroinflammatory response of tri-cultures to that of primary neuron-astrocyte co-cultures to a suite of known TLR agonists. We demonstrate that microglia are required for observation of neurotoxic neuroinflammatory responses, such as increased cell death and apoptosis, in response to TLR2, 3, 4, and 7/8 activation. Additionally, we show that following TLR3 agonist treatment, microglia and astrocytes play opposing roles in the neuroinflammatory response, and that the observed response is dictated by the degree of TLR3 activation. Overall, we demonstrate that microglia play a significant role in the neuroinflammatory response to TLR activation in vitro and, hence, the tri-culture has the potential to serve as a screening platform that better replicates the in vivo responses. |
| first_indexed | 2024-03-10T00:40:45Z |
| format | Article |
| id | doaj.art-d8c30d0022f74b7daadb28eaa4df523f |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-10T00:40:45Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomedicines |
| spelling | doaj.art-d8c30d0022f74b7daadb28eaa4df523f2023-11-23T15:09:27ZengMDPI AGBiomedicines2227-90592022-08-01109212210.3390/biomedicines10092122Primary Cortical Cell Tri-Culture-Based Screening of Neuroinflammatory Response in Toll-like Receptor ActivationNoah Goshi0Hyehyun Kim1Erkin Seker2Department of Biomedical Engineering, University of California—Davis, Davis, CA 95616, USADepartment of Biomedical Engineering, University of California—Davis, Davis, CA 95616, USADepartment of Electrical and Computer Engineering, University of California—Davis, Davis, CA 95616, USAThe activation of toll-like receptors (TLRs) in the central nervous system (CNS) can lead to neuroinflammation and contribute to many neurological disorders, including autoimmune diseases. Cell culture models are powerful tools for studying specific molecular and cellular mechanisms that contribute to these disease states and identifying potential therapeutics. However, most cell culture models have limitations in capturing biologically relevant phenomena, due in part to the non-inclusion of necessary cell types. Neurons, astrocytes, and microglia (critical cell types that play a role in neuroinflammation) all express at least a subset of TLRs. However, the response of each of these cell types to various TLR activation, along with their relative contribution to neuroinflammatory processes, is far from clear. In this study, we demonstrate the screening capabilities of a primary cortical cell tri-culture of neuron, astrocyte, and microglia from neonatal rats. Specifically, we compare the neuroinflammatory response of tri-cultures to that of primary neuron-astrocyte co-cultures to a suite of known TLR agonists. We demonstrate that microglia are required for observation of neurotoxic neuroinflammatory responses, such as increased cell death and apoptosis, in response to TLR2, 3, 4, and 7/8 activation. Additionally, we show that following TLR3 agonist treatment, microglia and astrocytes play opposing roles in the neuroinflammatory response, and that the observed response is dictated by the degree of TLR3 activation. Overall, we demonstrate that microglia play a significant role in the neuroinflammatory response to TLR activation in vitro and, hence, the tri-culture has the potential to serve as a screening platform that better replicates the in vivo responses.https://www.mdpi.com/2227-9059/10/9/2122microglianeuroinflammationin vitro modelmorphologytoll-like receptorCNS |
| spellingShingle | Noah Goshi Hyehyun Kim Erkin Seker Primary Cortical Cell Tri-Culture-Based Screening of Neuroinflammatory Response in Toll-like Receptor Activation Biomedicines microglia neuroinflammation in vitro model morphology toll-like receptor CNS |
| title | Primary Cortical Cell Tri-Culture-Based Screening of Neuroinflammatory Response in Toll-like Receptor Activation |
| title_full | Primary Cortical Cell Tri-Culture-Based Screening of Neuroinflammatory Response in Toll-like Receptor Activation |
| title_fullStr | Primary Cortical Cell Tri-Culture-Based Screening of Neuroinflammatory Response in Toll-like Receptor Activation |
| title_full_unstemmed | Primary Cortical Cell Tri-Culture-Based Screening of Neuroinflammatory Response in Toll-like Receptor Activation |
| title_short | Primary Cortical Cell Tri-Culture-Based Screening of Neuroinflammatory Response in Toll-like Receptor Activation |
| title_sort | primary cortical cell tri culture based screening of neuroinflammatory response in toll like receptor activation |
| topic | microglia neuroinflammation in vitro model morphology toll-like receptor CNS |
| url | https://www.mdpi.com/2227-9059/10/9/2122 |
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