etachment and the formation of spheroids under microgravity conditions can be observed with various types of intrinsically adherent human cells. In particular, for cancer cells this process mimics metastasis and may provide insights into cancer biology and progression that can be used to identify new drug/target combinations for future therapies. By using the synthetic glucocorticoid dexamethasone (DEX), we were able to suppress spheroid formation in a culture of follicular thyroid cancer (FTC)-133 cells that were exposed to altered gravity conditions on a random positioning machine. DEX inhibited the growth of three-dimensional cell aggregates in a dose-dependent manner. In the first approach, we analyzed the expression of several factors that are known to be involved in

Endothelial cells play a crucial role in the pathogenesis of many diseases and are highly sensitive to low gravity conditions. Using a three-dimensional random positioning machine (clinostat) we investigated effects of simulated weightlessness on the human EA.hy926 cell line (4, 12, 24, 48 and 72 h) and addressed the impact of exposure to VEGF (10 ng/ml). Simulated microgravity resulted in an increase in extracellular matrix proteins (ECMP) and altered cytoskeletal components such as microtubules (alpha-tubulin) and intermediate filaments (cytokeratin). Within the initial 4 h, both simulated microgravity and VEGF, alone, enhanced the expression of ECMP (collagen type I, fibronectin, osteopontin, laminin) and flk-1 protein.

With the expansion of commercial spaceflight and space exploration, the microgravity environment provides unparalleled opportunities to fight against challenging diseases. Here, we investigate the imp

Studying mammalian development is crucial for understanding the basis of developmental defects and for devising protocols for inducedpluripotent stem cell (iPSC) in vitro differentiation which can be used for cell transplantation. The inaccessibility of the advanced natural mammalian embryo due to its intimate association with the maternal uterus, that constitutes a “black box”,

Abstract The mammalian body plan is established shortly after the embryo implants into the maternal uterus, and our understanding of post-implantation developmental processes remains limited. Although pre- and peri-implantation mouse embryos are routinely cultured in vitro1,2, approaches for the robust culture of post-implantation embryos from egg cylinder stages until advanced organogenesis remain to be established. Here we present highly effective platforms for the ex utero culture of post-implantation mouse embryos, which enable the appropriate development of embryos from before gastrulation (embryonic day (E) 5.5) until the hindlimb formation stage (E11). Late gastrulating embryos (E7.5) are grown in three-dimensional rotating bottles, whereas extended culture from pre-gastrulation stages (E5.5 or E6.5) requires a combination of static and rotating bottle culture platforms. Histological, molecular and single-cell RNA sequencing analyses confirm that the ex utero cultured embryos recapitulate in utero development precisely. This culture system is amenable to the introduction of a variety of embryonic perturbations and micro-manipulations, the results of which can be followed ex utero for up to six days. The establishment of a system for robustly growing normal mouse embryos ex utero from pre-gastrulation to advanced organogenesis represents a valuable tool for investigating embryogenesis, as it eliminates the uterine barrier and allows researchers to mechanistically interrogate post-implantation morphogenesis and artificial embryogenesis in mammals.

Background: Tissue engineering is a strategy of cartilage regeneration, but scaffolds, required for 3D growth of chondrocytes, are still a problem. Methods: Searching for possibilities to improve scaffold-free engineering of cartilage, we characterized human chondrocytes incubated on a random positioning machine (RPM) to simulate microgravity (mu g). Results: When cultured in simulated mu g, human chondrocytes start forming 3D cell assemblies within 5 days. After 24h, we could not detect caspase-3, Fas, p53 or Bcl-2 proteins in these cells, Annexin V flow cytometry, however, revealed 18% of apoptotic chondrocytes in 1g cultures but only 10% on the RPM. Both rates of apoptosis were not changed, when vascular endothelial growth factor (VEGF) or basic fibroblast growth factor (bFGF) was added. 24 h, simulated microgravity also had significantly decreased collagen type I and X, but did not change collagen type IV and laminin, while collagen type II, chondroitin sulfate and aggrecan were elevated as compared with 1g controls. The production of collagen type II/X, chondroitin sulfate and aggrecan was modified, when external bFGF or VEGF had been applied. Conclusion: Chondrocytes exposed to simulated mu g seem to change their extracellular matrix production behavior, while they rearrange their cytoskeletal proteins prior to forming 3D aggregates. Copyright (C) 2010 S. Karger AG, Basel

In vitro cultured stem cells with distinct developmental capacities can contribute to embryonic or extraembryonictissues after microinjection into pre-implantation mammalian embryos. However, whether

The mammalian body plan is established shortly after the embryo implants into the maternal uterus, and our understanding of post-implantation developmental processes remains limited. Although pre- and peri-implantation mouse embryos are routinely cultured in vitro1,2, approaches for the robust culture of post-implantation embryos from egg cylinder stages until advanced organogenesis remain to be established. Here we present highly effective platforms for the ex utero culture of post-implantation mouse embryos, which enable the appropriate development of embryos from before gastrulation (embryonic day (E) 5.5) until the hindlimb formation stage (E11). Late gastrulating embryos (E7.5) are grown in threedimensional rotating bottles, whereas extended culture from pre-gastrulation stages (E5.5 or E6.5) requires a combination of static and rotating bottle culture platforms.

With the commercialization of spaceflight and the exploration of space, it is important to understand the changes occurring in human cells exposed to real microgravity (r-mu g) conditions. We examined the influence of r-mu g, simulated microgravity (s-mu g, incubator random positioning machine (iRPM)), hypergravity (hyper-g), and vibration (VIB) on triple-negative breast cancer (TNBC) cells (MDA-MB-231 cell line) with the aim to study early changes in the gene expression of factors associated with cell adhesion, apoptosis,

​The effect of clinostat-simulated microgravity on SP-2/0 and 1D6 hybridoma cells was studied. Clinorotation during 4–5 days at 1.5 rounds per minute decreased dramatically their proliferating capacity: the rotated cells divided less than once while control cells performed 4–5 divisions. They decreased the non-specific adhesion to tissue culture plastic, but increased the number of cell-to-cell contacts. Such phenomenological changes were accompanied with the alterations in pericellular glycosaminoglycans:

宇宙环境对航天员的身体健康带来了一些不良影响。目前,关于空间环境,尤其是微重力对宇航员中枢神经系统影响的研究层出不穷。但是少有将不同脑区的微重力效应进行比较的研究进展。不同脑区在细胞组成、组织结构、功能作用等方面都存在差异,所以不同脑区对微重力的响应是否相同的问题也需要解决。研究地面模拟微重力效应对不同脑区组织形态学和凋亡的影响,是研究微重力效应的重要一环。

​Space travel is a major challenge for human beings. Especially, the mechanisms through which space conditions might alter animal development have been questioned for a long time. The two major physical stress factors that are of relevance in this context are space radiation and weightlessness. While it has been extensively shown that high doses of ionizing radiation induce deleterious effects on embryonic development