Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning

Frontiers in Cell and Developmental Biology

Cell and Developmental Biology

Christina James¹  Kenyi Saito-Diaz¹  Archie Jayesh Patel¹  Nadja Zeltner² 

[1] Center for Molecular Medicine, University of Georgia, Athens, GA, United States;Center for Molecular Medicine, University of Georgia, Athens, GA, United States;Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, United States;Department of Cellular Biology, University of Georgia, Athens, GA, United States;

关键词: peripheral nervous system (PNS);    human pluripotent stem cell (hPSC);    sensory neurons;    nociceptor;    mechanoreceptor;    proprioceptor;    immunopanning;   

DOI  :  10.3389/fcell.2023.1101423

 received in 2022-11-17, accepted in 2023-04-12,  发布年份 2023

来源: Frontiers

【 摘 要 】

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.

【 授权许可】

Unknown   
Copyright © 2023 Saito-Diaz, James, Patel and Zeltner.