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

IF 4.4 1区 物理与天体物理 Q1 MULTIDISCIPLINARY SCIENCES
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
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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.

表面张力使诱导多能干细胞能在太空飞行期间通过市售硬件进行培养。
低地球轨道(LEO)已成为评估微重力下干细胞特性改变的独特环境。由于私人太空飞行的进步,越来越多的人可以利用低地轨道进行研究和开发。Axiom任务2(Ax-2)作为第二次全私人宇航员任务被发射到国际空间站(ISS)。在SOX2启动子下表达绿色荧光蛋白(GFP)的冷冻人类诱导多能干细胞(hiPSC)以及由SOX2-GFP hiPSC分化而成的成纤维细胞被送往国际空间站。宇航员随后解冻了这两种细胞,并将其播种到市售的96孔板中,这些孔板的表面张力可减少液体流出单个孔,结果表明,hiPSC或hiPSC衍生的成纤维细胞既可以悬浮存活,也可以附着在Matrigel基底上存活。此外,这两种细胞类型都能转染表达红色荧光蛋白(RFP)的质粒。我们证明,hiPSC 和 hiPSC-成纤维细胞可以在微重力环境下通过现成的商用细胞培养硬件解冻,可以结合成三维球体或在 Matrigel 中粘附生长,还可以用 DNA 进行转染。这为未来的太空生物制造实验奠定了基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
npj Microgravity
npj Microgravity Physics and Astronomy-Physics and Astronomy (miscellaneous)
CiteScore
7.30
自引率
7.80%
发文量
50
审稿时长
9 weeks
期刊介绍: A new open access, online-only, multidisciplinary research journal, npj Microgravity is dedicated to publishing the most important scientific advances in the life sciences, physical sciences, and engineering fields that are facilitated by spaceflight and analogue platforms.
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