Literature DB >> 32455868

A Temperature-Controlled Patch Clamp Platform Demonstrated on Jurkat T Lymphocytes and Human Induced Pluripotent Stem Cell-Derived Neurons.

Jann Harberts1, Max Kusch1, John O'Sullivan1,2, Robert Zierold1, Robert H Blick1,3.   

Abstract

Though patch clamping at room temperature is a widely disseminated standard procedure in the electrophysiological community, it does not represent the biological system in mammals at around 37 °C. In order to better mimic the natural environment in electrophysiological studies, we present a custom-built, temperature-controlled patch clamp platform for upright microscopes, which can easily be adapted to any upright patch clamp setup independently, whether commercially available or home built. Our setup can both cool and heat the platform having only small temperature variations of less than 0.5 °C. We demonstrate our setup with patch clamp measurements at 36 °C on Jurkat T lymphocytes and human induced pluripotent stem cell-derived neurons. Passive membrane parameters and characteristic electrophysiological properties, such as the gating properties of voltage-gated ion channels and the firing of action potentials, are compared to measurements at room temperature. We observe that many processes that are not explicitly considered as temperature dependent show changes with temperature. Thus, we believe in the need of a temperature control in patch clamp measurements if improved physiological conditions are required. Furthermore, we advise researchers to only compare electrophysiological results directly that have been measured at similar temperatures since small variations in cellular properties might be caused by temperature alterations.

Entities:  

Keywords:  Jurkat cells; electrophysiology; human induced pluripotent stem cell-derived neurons; patch clamping; temperature control

Year:  2020        PMID: 32455868     DOI: 10.3390/bioengineering7020046

Source DB:  PubMed          Journal:  Bioengineering (Basel)        ISSN: 2306-5354


  2 in total

1.  Robust neuronal differentiation of human iPSC-derived neural progenitor cells cultured on densely-spaced spiky silicon nanowire arrays.

Authors:  Jann Harberts; Malte Siegmund; Matteo Schnelle; Ting Zhang; Yakui Lei; Linwei Yu; Robert Zierold; Robert H Blick
Journal:  Sci Rep       Date:  2021-09-22       Impact factor: 4.379

2.  Versatile bipolar temperature controller for custom in vitro applications.

Authors:  Sabrina Asteriti; Lorenzo Cangiano
Journal:  HardwareX       Date:  2020-10
  2 in total

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