Spheroid formation of human thyroid cancer cells in an automated culturing system during the Shenzhou-8 Space mission.

Human follicular thyroid cancer cells were cultured in Space to investigate the impact of microgravity on 3D growth. For this purpose, we designed and constructed a cell container that can endure enhanced physical forces, is connected to fluid storage chambers, performs media changes and cell harvesting automatically and supports cell viability. The container consists of a cell suspension chamber, two reserve tanks for medium and fixative and a pump for fluid exchange. The selected materials proved durable, non-cytotoxic, and did not inactivate RNAlater. This container was operated automatically during the unmanned Shenzhou-8 Space mission. FTC-133 human follicular thyroid cancer cells were cultured in Space for 10 days. Culture medium was exchanged after 5 days in Space and the cells were fixed after 10 days. The experiment revealed a scaffold-free formation of extraordinary large three-dimensional aggregates by thyroid cancer cells with altered expression of EGF and CTGF genes under real microgravity.

[1]  D. Grimm,et al.  Establishment and characterization of the follicular thyroid carcinoma cell line ML-1 , 2000, Journal of Molecular Medicine.

[2]  A. Cogoli,et al.  Neocartilage formation in 1 g, simulated, and microgravity environments: implications for tissue engineering. , 2010, Tissue engineering. Part A.

[3]  Zhen-ning Wang,et al.  Downregulation of connective tissue growth factor inhibits the growth and invasion of gastric cancer cells and attenuates peritoneal dissemination , 2011, Molecular Cancer.

[4]  A. Frilling,et al.  Growth regulation of normal thyroids and thyroid tumors in man. , 1990, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.

[5]  Augusto Cogoli,et al.  Effects of basic fibroblast growth factor on endothelial cells under conditions of simulated microgravity , 2008, Journal of cellular biochemistry.

[6]  Roy S Herbst,et al.  Review of epidermal growth factor receptor biology. , 2004, International journal of radiation oncology, biology, physics.

[7]  G. Schulze-Tanzil,et al.  Simulated microgravity alters differentiation and increases apoptosis in human follicular thyroid carcinoma cells , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[8]  M. Braun,et al.  The Impact of Altered Gravity and Vibration on Endothelial Cells During a Parabolic Flight , 2013, Cellular Physiology and Biochemistry.

[9]  A. Lendlein,et al.  Cytocompatibility testing of cell culture modules fabricated from specific candidate biomaterials using injection molding. , 2010, Journal of biotechnology.

[10]  Lei Cui,et al.  CTGF is overexpressed in papillary thyroid carcinoma and promotes the growth of papillary thyroid cancer cells , 2011, Tumor Biology.

[11]  G R Taylor,et al.  Overview of spaceflight immunology studies , 1993, Journal of leukocyte biology.

[12]  Min Xu,et al.  Simulated microgravity using the Random Positioning Machine inhibits differentiation and alters gene expression profiles of 2T3 preosteoblasts. , 2005, American journal of physiology. Cell physiology.

[13]  H. Fine,et al.  Effect of brain- and tumor-derived connective tissue growth factor on glioma invasion. , 2011, Journal of the National Cancer Institute.

[14]  D. Grimm,et al.  The effects of weightlessness on the human organism and mammalian cells. , 2011, Current molecular medicine.

[15]  Kriss Westphal,et al.  A delayed type of three-dimensional growth of human endothelial cells under simulated weightlessness. , 2009, Tissue engineering. Part A.

[16]  J. Folkman.  Tumor angiogenesis: therapeutic implications. , 1971, The New England journal of medicine.

[17]  Kriss Westphal,et al.  Characterization of Human Chondrocytes Exposed to Simulated Microgravity , 2010, Cellular Physiology and Biochemistry.

[18]  W. Mueller‐Klieser,et al.  Multicellular spheroids , 2004, Journal of Cancer Research and Clinical Oncology.

[19]  M. Shakibaei,et al.  Mechanisms of apoptosis after ischemia and reperfusion: Role of the renin-angiotensin system , 2006, Apoptosis.

[20]  K. Honn,et al.  12-Lipoxygenase in Lewis lung carcinoma cells: molecular identity, intracellular distribution of activity and protein, and Ca(2+)-dependent translocation from cytosol to membranes. , 1995, Prostaglandins.

[21]  D. Grimm,et al.  Human follicular and papillary thyroid carcinoma cells interact differently with human venous endothelial cells. , 1995, Thyroid : official journal of the American Thyroid Association.

[22]  D. Grimm,et al.  Proteome Analysis of Thyroid Cancer Cells After Long-Term Exposure to a Random Positioning Machine , 2011 .

[23]  P. Meltzer,et al.  Vascular channel formation by human melanoma cells in vivo and in vitro: vasculogenic mimicry. , 1999, The American journal of pathology.

[24]  M. Shakibaei,et al.  Simulated weightlessness changes the cytoskeleton and extracellular matrix proteins in papillary thyroid carcinoma cells , 2006, Cell and Tissue Research.

[25]  Felice Strollo,et al.  Chapter 4 Hormonal Changes in Humans During Spaceflight , 1999 .

[26]  D. Grimm,et al.  Interleukin-6 Expression under Gravitational Stress Due to Vibration and Hypergravity in Follicular Thyroid Cancer Cells , 2013, PloS one.

[27]  Robert J.P. Williams.  ALUMINIUM AND BIOLOGICAL SYSTEMS : AN INTRODUCTION , 1996 .

[28]  G. Gronowicz,et al.  The in vitro response of human osteoblasts to polyetheretherketone (PEEK) substrates compared to commercially pure titanium. , 2008, Biomaterials.

[29]  D. Grimm,et al.  Radiolabeled annexin V for imaging apoptosis in radiated human follicular thyroid carcinomas--is an individualized protocol necessary? , 2009, Nuclear medicine and biology.

[30]  Pankaj Gupta,et al.  SimBOX: a scalable architecture for aggregate distributed command and control of spaceport and service constellation , 2004, SPIE Defense + Commercial Sensing.

[31]  K. Furuya,et al.  Temperature dependency of calcium responses in mammary tumour cells , 1997, Cell biochemistry and function.

[32]  R. Wildgruber,et al.  Application of free‐flow IEF to identify protein candidates changing under microgravity conditions , 2010, Proteomics.

[33]  Jack J. W. A. van Loon,et al.  Some history and use of the random positioning machine, RPM, in gravity related research , 2007 .

[34]  Jens Hauslage,et al.  Short‐term weightlessness produced by parabolic flight maneuvers altered gene expression patterns in human endothelial cells , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[35]  V V Polyakov,et al.  The alteration of human sleep and circadian rhythms during spaceflight , 1997, Journal of sleep research.

[36]  C. D. Kramer,et al.  The challenge of designing biomedical equipment during human research for long duration low-gravity NASA missions , 1997, Proceedings of the 1997 16 Southern Biomedical Engineering Conference.

[37]  A L Goldberger,et al.  An episode of ventricular tachycardia during long-duration spaceflight. , 1998, The American journal of cardiology.

[38]  Yoshio Masuda,et al.  Evaluation of the three-dimensional clinostat as a simulator of weightlessness , 1997, Planta.

[39]  J. Redpath,et al.  Reduced temperature (22 degrees C) results in enhancement of cell killing and neoplastic transformation in noncycling HeLa x skin fibroblast human hybrid cells irradiated with low-dose-rate gamma radiation. , 1995, Radiation research.

[40]  R. Wildgruber,et al.  A proteomic approach to analysing spheroid formation of two human thyroid cell lines cultured on a random positioning machine , 2011, Proteomics.

[41]  Jens Hauslage,et al.  Ground-based facilities for simulation of microgravity: organism-specific recommendations for their use, and recommended terminology. , 2013, Astrobiology.

[42]  K M Baldwin,et al.  Effect of spaceflight on the functional, biochemical, and metabolic properties of skeletal muscle. , 1996, Medicine and science in sports and exercise.

[43]  D. Grimm,et al.  Expression of vascular endothelial growth factor and receptor tyrosine kinases in cardiac ischemia/reperfusion injury. , 2007, Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology.

[44]  Ken Saito,et al.  Vasculogenic mimicry and pseudo‐comedo formation in breast cancer , 2002, International journal of cancer.

[45]  H. Sampson,et al.  Standardization and performance evaluation of mononuclear cell cytokine secretion assays in a multicenter study , 2006, BMC Immunology.

[46]  F. Strollo.  Hormonal changes in humans during spaceflight. , 1999, Advances in space biology and medicine.

[47]  Ruth Hemmersbach,et al.  Gravity‐sensitive signaling drives 3‐dimensional formation of multicellular thyroid cancer spheroids , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[48]  Ronald J. White,et al.  Humans in space , 2001, Nature.

[49]  A. Cogoli,et al.  Signal transduction in T lymphocytes--a comparison of the data from space, the free fall machine and the random positioning machine. , 1999, Advances in space research : the official journal of the Committee on Space Research.

[50]  G. Polyzois,et al.  Physicomechanical and cytotoxic properties of room temperature vulcanizing silicone prosthetic elastomers. , 1998, Acta odontologica Scandinavica.

[51]  R. Wildgruber,et al.  Metabolic enzyme diversity in different human thyroid cell lines and their sensitivity to gravitational forces , 2012, Proteomics.

[52]  R. Einspanier,et al.  Differential Gene Regulation under Altered Gravity Conditions in Follicular Thyroid Cancer Cells: Relationship between the Extracellular Matrix and the Cytoskeleton , 2011, Cellular Physiology and Biochemistry.

[53]  D. Puleo,et al.  Acute toxicity of metal ions in cultures of osteogenic cells derived from bone marrow stromal cells. , 1995, Journal of applied biomaterials : an official journal of the Society for Biomaterials.