Journal article
A More Rapid Method for Culturing LUHMES-Derived Neurons Provides Greater Cell Numbers and Facilitates Studies of Multiple Viruses
Viruses, Vol.17(7), 1001
07/16/2025
DOI: 10.3390/v17071001
PMCID: PMC12299948
PMID: 40733616
Abstract
The ability to study mature neuronal cells ex vivo is complicated by their non-dividing nature and difficulty in obtaining large numbers of primary cells from organisms. Thus, numerous transformed progenitor models have been developed that can be routinely cultured, then scaled, and differentiated to mature neurons. In this paper, we present a new method for differentiating one such model, the Lund human mesencephalic (LUHMES) dopaminergic neurons. This method is two days faster than some established protocols, results in nearly five times greater numbers of mature neurons, and involves fewer handling steps that could introduce technical variability. Moreover, it overcomes the problem of cell aggregate formation that commonly impedes high-resolution imaging, cell dissociation, and downstream analysis. While recently established for herpes simplex virus type 1, we demonstrate that LUHMES neurons can facilitate studies of other herpesviruses, as well as RNA viruses associated with childhood encephalitis and hemorrhagic fever. This protocol provides an improvement in the generation of large-scale neuronal cultures, which may be readily applicable to other neuronal 2D cell culture models and provides a system for studying neurotrophic viruses. We named this method the Streamlined Protocol for Enhanced Expansion and Differentiation Yield, or SPEEDY, method.
Details
- Title: Subtitle
- A More Rapid Method for Culturing LUHMES-Derived Neurons Provides Greater Cell Numbers and Facilitates Studies of Multiple Viruses
- Creators
- Adam Whisnant - University of IowaStephanie Clark - University of IowaJosé Aguilar-Briseño - University of Iowa, Microbiology and ImmunologyLorellin Durnell - University of IowaArnhild Grothey - University of WürzburgAnn Miller - University of IowaSteven Varga - University of IowaJeffery Meier - University of IowaCharles Grose - University of IowaPatrick Sinn - University of IowaJessica Tucker - University of IowaCaroline Friedel - Ludwig-Maximilians-Universität MünchenWendy Maury - University of IowaDavid Price - University of IowaDölken Lars - University of Würzburg
- Resource Type
- Journal article
- Publication Details
- Viruses, Vol.17(7), 1001
- DOI
- 10.3390/v17071001
- PMID
- 40733616
- PMCID
- PMC12299948
- NLM abbreviation
- Viruses
- ISSN
- 1999-4915
- eISSN
- 1999-4915
- Publisher
- MDPI AG
- Grant note
- European Research CouncilNational Institutes of Health: GM126908 Deutsche Forschungsgemeinschaft (DFG, German Research Foundation): FOR5200 DEEP-DV, 443644894, FR 2938/11-1 Department of Veterans Affairs Merit Award: BX004434 NIH: GM126908
This research was funded by the European Research Council ERC-2021-CoG 101041177-DecipherHSV to LD and the National Institutes of Health GM126908 to DHP. CCF was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) within the framework of the Research Unit FOR5200 DEEP-DV (443644894) project FR 2938/11-1. JLM was supported by the Department of Veterans Affairs Merit Award BX004434. The APC was funded by NIH GM126908.
- Language
- English
- Date published
- 07/16/2025
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Infectious Diseases; Stead Family Department of Pediatrics; Epidemiology; Biochemistry and Molecular Biology; Infectious Disease (Pediatrics); Internal Medicine
- Record Identifier
- 9984865315902771
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