Krüger Marcus, Pietsch Jessica, Bauer Johann, Kopp Sascha, Carvalho Daniel T O, Baatout Sarah, Moreels Marjan, Melnik Daniela, Wehland Markus, Egli Marcel, Jayashree Sahana, Kobberø Sara Dam, Corydon Thomas J, Nebuloni Stefano, Gass Samuel, Evert Matthias, Infanger Manfred, Grimm Daniel
Clinic for Plastic, Aesthetic and Hand Surgery, Otto von Guericke University Magdeburg, Magdeburg, Germany.
Max Planck Institute of Biochemistry, Martinsried, Germany.
Cell Physiol Biochem. 2019;52(5):1039-1060. doi: 10.33594/000000071.
BACKGROUND/AIMS: Endothelial cells exposed to the Random Positioning Machine (RPM) reveal three different phenotypes. They grow as a two-dimensional monolayer and form three-dimensional (3D) structures such as spheroids and tubular constructs. As part of the ESA-SPHEROIDS project we want to understand how endothelial cells (ECs) react and adapt to long-term microgravity.
During a spaceflight to the International Space Station (ISS) and a subsequent stay onboard, human ECs (EA.hy926 cell line) were cultured for 12 days in real microgravity inside an automatic flight hardware, specially designed for use in space. ECs were cultivated in the absence or presence of vascular endothelial growth factor, which had demonstrated a cell-protective effect on ECs exposed to an RPM simulating microgravity. After cell fixation in space and return of the samples, we examined cell morphology and analyzed supernatants by Multianalyte Profiling technology.
The fixed samples comprised 3D multicellular spheroids and tube-like structures in addition to monolayer cells, which are exclusively observed during growth under Earth gravity (1g). Within the 3D aggregates we detected enhanced collagen and laminin. The supernatant analysis unveiled alterations in secretion of several growth factors, cytokines, and extracellular matrix components as compared to cells cultivated at 1g or on the RPM. This confirmed an influence of gravity on interacting key proteins and genes and demonstrated a flight hardware impact on the endothelial secretome.
Since formation of tube-like aggregates was observed only on the RPM and during spaceflight, we conclude that microgravity may be the major cause for ECs' 3D aggregation.
背景/目的：暴露于随机定位机（RPM）的内皮细胞呈现出三种不同的表型。它们以二维单层形式生长，并形成三维（3D）结构，如球体和管状结构。作为欧洲航天局 - 球体项目的一部分，我们希望了解内皮细胞（ECs）如何对长期微重力做出反应和适应。
在前往国际空间站（ISS）的太空飞行及随后在空间站的停留期间，人类内皮细胞（EA.hy926细胞系）在专门设计用于太空的自动飞行硬件内的真实微重力环境中培养12天。内皮细胞在有无血管内皮生长因子的情况下进行培养，该因子已证明对暴露于模拟微重力的RPM的内皮细胞具有细胞保护作用。在太空中进行细胞固定并将样品返回后，我们检查了细胞形态，并通过多分析物谱技术分析了上清液。
固定后的样品除了单层细胞外，还包括3D多细胞球体和管状结构，而单层细胞仅在地球重力（1g）下生长时观察到。在3D聚集体中，我们检测到胶原蛋白和层粘连蛋白增加。上清液分析显示，与在1g或RPM上培养的细胞相比，几种生长因子、细胞因子和细胞外基质成分的分泌发生了变化。这证实了重力对相互作用的关键蛋白质和基因的影响，并证明了飞行硬件对内皮分泌组的影响。
由于仅在RPM上和太空飞行期间观察到管状聚集体的形成，我们得出结论，微重力可能是内皮细胞3D聚集的主要原因。
