Electroporation outcomes in human Jurkat cells are affected by exposure to simulated microgravity

Matthew T. Conway; Anna V. Sedelnikova; Edward A. Sander; Kristan S. Worthington; Zachary A. Steelman

doi:10.21203/rs.3.rs-10896773/v1

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Electroporation outcomes in human Jurkat cells are affected by exposure to simulated microgravity

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Electroporation outcomes in human Jurkat cells are affected by exposure to simulated microgravity

Matthew T. Conway, Anna V. Sedelnikova, Edward A. Sander, Kristan S. Worthington and Zachary A. Steelman

Research Square

American Journal Experts

09/15/2026

DOI: 10.21203/rs.3.rs-10896773/v1

PMCID: PMC13596624

PMID: 42780173

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https://doi.org/10.21203/rs.3.rs-10896773/v1View

Preprint (Author's original) This preprint has not been evaluated by subject experts through peer review. Preprints may undergo extensive changes and/or become peer-reviewed journal articles. Open Access

Abstract

Electroporation (EP) is commonly used to permeabilize cell membranes to deliver nucleic acids and other biomolecules. EP-based methodologies have the potential to support in-space biomanufacturing; however, it is currently unclear whether cellular adaptations to microgravity influence EP outcomes. To investigate this, human T-lymphocyte (Jurkat) cells were exposed to simulated microgravity (SµG) in a rotating wall vessel (RWV) bioreactor prior to EP to deliver exogenous fluorescent molecules or plasmid DNA. We observed that two hours of SµG exposure led to a 20% decrease in fluorophore uptake and a 31% decrease in transfection efficiency compared to standard gravity controls for several voltages. To explore underlying mechanisms, the effects of SµG on actin organization were assessed using a phalloidin stain, which revealed increased cell area and cortical actin thickness following SµG. Actin targeting drugs jasplakinolide and latrunculin B were used to promote or destabilize F-actin, respectively, and modulated cell area and cortical actin thickness. Promoting F-actin also led to decreased transfection efficiency, mimicking the effect of SµG, while destabilizing F-actin during SµG showed a small increase in transfection efficiency, slightly reversing the impact of SµG. Our results demonstrate that adaptations to microgravity exposure can influence transfection efficiency via cytoskeletal reorganization.

Details

Title: Subtitle: Electroporation outcomes in human Jurkat cells are affected by exposure to simulated microgravity
Creators: Matthew T. Conway - University of Iowa
Anna V. Sedelnikova - Science Applications International Corporation (United States)
Edward A. Sander - University of Iowa
Kristan S. Worthington - University of Iowa
Zachary A. Steelman - Mitchell Institute
Resource Type: Preprint
Publication Details: Research Square
DOI: 10.21203/rs.3.rs-10896773/v1
PMID: 42780173
PMCID: PMC13596624
eISSN: 2693-5015
Publisher: American Journal Experts
Language: English
Date posted: 09/15/2026
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Iowa Neuroscience Institute; Orthopedics and Rehabilitation; Chemical and Biochemical Engineering
Record Identifier: 9985231452802771

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