Surface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflight

Abstract Low Earth Orbit (LEO) has emerged as a unique environment for evaluating altered stem cell properties in microgravity. LEO has become increasingly accessible for research and development due to progress in private spaceflight. Axiom Mission 2 (Ax-2) was launched as the second all-private as...

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Main Authors: Maedeh Mozneb, Madelyn Arzt, Pinar Mesci, Dylan M. N. Martin, Stephany Pohlman, George Lawless, Shankini Doraisingam, Sultan Al Neyadi, Rayyanah Barnawi, Ali Al Qarni, Peggy A. Whitson, John Shoffner, Jana Stoudemire, Stefanie Countryman, Clive N. Svendsen, Arun Sharma
Format: Article
Language:English
Published: Nature Portfolio 2024-10-01
Series:npj Microgravity
Online Access:https://doi.org/10.1038/s41526-024-00435-y
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author Maedeh Mozneb
Madelyn Arzt
Pinar Mesci
Dylan M. N. Martin
Stephany Pohlman
George Lawless
Shankini Doraisingam
Sultan Al Neyadi
Rayyanah Barnawi
Ali Al Qarni
Peggy A. Whitson
John Shoffner
Jana Stoudemire
Stefanie Countryman
Clive N. Svendsen
Arun Sharma
author_facet Maedeh Mozneb
Madelyn Arzt
Pinar Mesci
Dylan M. N. Martin
Stephany Pohlman
George Lawless
Shankini Doraisingam
Sultan Al Neyadi
Rayyanah Barnawi
Ali Al Qarni
Peggy A. Whitson
John Shoffner
Jana Stoudemire
Stefanie Countryman
Clive N. Svendsen
Arun Sharma
author_sort Maedeh Mozneb
collection DOAJ
description Abstract Low Earth Orbit (LEO) has emerged as a unique environment for evaluating altered stem cell properties in microgravity. LEO has become increasingly accessible for research and development due to progress in private spaceflight. Axiom Mission 2 (Ax-2) was launched as the second all-private astronaut mission to the International Space Station (ISS). Frozen human induced pluripotent stem cells (hiPSCs) expressing green fluorescent protein (GFP) under the SOX2 promoter, as well as fibroblasts differentiated from SOX2-GFP hiPSCs, were sent to the ISS. Astronauts then thawed and seeded both cell types into commercially available 96-well plates, which provided surface tension that reduced fluid movement out of individual wells and showed that hiPSCs or hiPSC-derived fibroblasts could survive either in suspension or attached to a Matrigel substrate. Furthermore, both cell types could be transfected with red fluorescent protein (RFP)-expressing plasmid. We demonstrate that hiPSCs and hiPSC-fibroblasts can be thawed in microgravity in off-the-shelf, commercially-available cell culture hardware, can associate into 3D spheroids or grow adherently in Matrigel, and can be transfected with DNA. This lays the groundwork for future biomanufacturing experiments in space.
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series npj Microgravity
spelling doaj-art-6041df6b54fe481daba4e2748c8fe5742025-08-20T02:17:48ZengNature Portfolionpj Microgravity2373-80652024-10-0110111010.1038/s41526-024-00435-ySurface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflightMaedeh Mozneb0Madelyn Arzt1Pinar Mesci2Dylan M. N. Martin3Stephany Pohlman4George Lawless5Shankini Doraisingam6Sultan Al Neyadi7Rayyanah Barnawi8Ali Al Qarni9Peggy A. Whitson10John Shoffner11Jana Stoudemire12Stefanie Countryman13Clive N. Svendsen14Arun Sharma15Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical CenterBoard of Governors Regenerative Medicine Institute, Cedars-Sinai Medical CenterAxiom Space, Inc.BioServe Space TechnologiesBoard of Governors Regenerative Medicine Institute, Cedars-Sinai Medical CenterBoard of Governors Regenerative Medicine Institute, Cedars-Sinai Medical CenterBioServe Space TechnologiesMohammed bin Rashid Space CentreAxiom Space, Inc.Axiom Space, Inc.Axiom Space, Inc.Axiom Space, Inc.Axiom Space, Inc.BioServe Space TechnologiesBoard of Governors Regenerative Medicine Institute, Cedars-Sinai Medical CenterBoard of Governors Regenerative Medicine Institute, Cedars-Sinai Medical CenterAbstract Low Earth Orbit (LEO) has emerged as a unique environment for evaluating altered stem cell properties in microgravity. LEO has become increasingly accessible for research and development due to progress in private spaceflight. Axiom Mission 2 (Ax-2) was launched as the second all-private astronaut mission to the International Space Station (ISS). Frozen human induced pluripotent stem cells (hiPSCs) expressing green fluorescent protein (GFP) under the SOX2 promoter, as well as fibroblasts differentiated from SOX2-GFP hiPSCs, were sent to the ISS. Astronauts then thawed and seeded both cell types into commercially available 96-well plates, which provided surface tension that reduced fluid movement out of individual wells and showed that hiPSCs or hiPSC-derived fibroblasts could survive either in suspension or attached to a Matrigel substrate. Furthermore, both cell types could be transfected with red fluorescent protein (RFP)-expressing plasmid. We demonstrate that hiPSCs and hiPSC-fibroblasts can be thawed in microgravity in off-the-shelf, commercially-available cell culture hardware, can associate into 3D spheroids or grow adherently in Matrigel, and can be transfected with DNA. This lays the groundwork for future biomanufacturing experiments in space.https://doi.org/10.1038/s41526-024-00435-y
spellingShingle Maedeh Mozneb
Madelyn Arzt
Pinar Mesci
Dylan M. N. Martin
Stephany Pohlman
George Lawless
Shankini Doraisingam
Sultan Al Neyadi
Rayyanah Barnawi
Ali Al Qarni
Peggy A. Whitson
John Shoffner
Jana Stoudemire
Stefanie Countryman
Clive N. Svendsen
Arun Sharma
Surface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflight
npj Microgravity
title Surface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflight
title_full Surface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflight
title_fullStr Surface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflight
title_full_unstemmed Surface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflight
title_short Surface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflight
title_sort surface tension enables induced pluripotent stem cell culture in commercially available hardware during spaceflight
url https://doi.org/10.1038/s41526-024-00435-y
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