Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning
Sensory neurons (SNs) detect a wide range of information from the body and the environment that is critical for homeostasis. There are three main subtypes of SNs: nociceptors, mechanoreceptors, and proprioceptors, which express different membrane proteins, such as TRKA, TRKB, or TRKC, respectively....
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2023.1101423/full |
| _version_ | 1827955045246697472 |
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| author | Kenyi Saito-Diaz Christina James Archie Jayesh Patel Nadja Zeltner Nadja Zeltner Nadja Zeltner |
| author_facet | Kenyi Saito-Diaz Christina James Archie Jayesh Patel Nadja Zeltner Nadja Zeltner Nadja Zeltner |
| author_sort | Kenyi Saito-Diaz |
| collection | DOAJ |
| description | Sensory neurons (SNs) detect a wide range of information from the body and the environment that is critical for homeostasis. There are three main subtypes of SNs: nociceptors, mechanoreceptors, and proprioceptors, which express different membrane proteins, such as TRKA, TRKB, or TRKC, respectively. Human pluripotent stem cell technology provides an ideal platform to study development and diseases of SNs, however there is not a viable method to isolate individual SN subtype for downstream analysis available. Here, we employ the method immunopanning to isolate each SN subtype. This method is very gentle and allows proper survival after the isolation. We use antibodies against TRKA, TRKB, and TRKC to isolate nociceptors, mechanoreceptors, and proprioceptors, respectively. We show that our cultures are enriched for each subtype and express their respective subtype markers. Furthermore, we show that the immunopanned SNs are electrically active and respond to specific stimuli. Thus, our method can be used to purify viable neuronal subtypes using respective membrane proteins for downstream studies. |
| first_indexed | 2024-04-09T14:39:45Z |
| format | Article |
| id | doaj.art-0b0f38f7fc83464288005ab616638cad |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-09T14:39:45Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-0b0f38f7fc83464288005ab616638cad2023-05-03T08:38:48ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2023-05-011110.3389/fcell.2023.11014231101423Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanningKenyi Saito-Diaz0Christina James1Archie Jayesh Patel2Nadja Zeltner3Nadja Zeltner4Nadja Zeltner5Center for Molecular Medicine, University of Georgia, Athens, GA, United StatesCenter for Molecular Medicine, University of Georgia, Athens, GA, United StatesCenter for Molecular Medicine, University of Georgia, Athens, GA, United StatesCenter for Molecular Medicine, University of Georgia, Athens, GA, United StatesDepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, United StatesDepartment of Cellular Biology, University of Georgia, Athens, GA, United StatesSensory neurons (SNs) detect a wide range of information from the body and the environment that is critical for homeostasis. There are three main subtypes of SNs: nociceptors, mechanoreceptors, and proprioceptors, which express different membrane proteins, such as TRKA, TRKB, or TRKC, respectively. Human pluripotent stem cell technology provides an ideal platform to study development and diseases of SNs, however there is not a viable method to isolate individual SN subtype for downstream analysis available. Here, we employ the method immunopanning to isolate each SN subtype. This method is very gentle and allows proper survival after the isolation. We use antibodies against TRKA, TRKB, and TRKC to isolate nociceptors, mechanoreceptors, and proprioceptors, respectively. We show that our cultures are enriched for each subtype and express their respective subtype markers. Furthermore, we show that the immunopanned SNs are electrically active and respond to specific stimuli. Thus, our method can be used to purify viable neuronal subtypes using respective membrane proteins for downstream studies.https://www.frontiersin.org/articles/10.3389/fcell.2023.1101423/fullperipheral nervous system (PNS)human pluripotent stem cell (hPSC)sensory neuronsnociceptormechanoreceptorproprioceptor |
| spellingShingle | Kenyi Saito-Diaz Christina James Archie Jayesh Patel Nadja Zeltner Nadja Zeltner Nadja Zeltner Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning Frontiers in Cell and Developmental Biology peripheral nervous system (PNS) human pluripotent stem cell (hPSC) sensory neurons nociceptor mechanoreceptor proprioceptor |
| title | Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning |
| title_full | Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning |
| title_fullStr | Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning |
| title_full_unstemmed | Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning |
| title_short | Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning |
| title_sort | isolation of human pluripotent stem cell derived sensory neuron subtypes by immunopanning |
| topic | peripheral nervous system (PNS) human pluripotent stem cell (hPSC) sensory neurons nociceptor mechanoreceptor proprioceptor |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2023.1101423/full |
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