Evaluating the effect of spaceflight on the host-pathogen interaction between human intestinal epithelial cells and Salmonella Typhimurium.

Spaceflight uniquely alters the physiology of both human cells and microbial pathogens, stimulating cellular and molecular changes directly relevant to infectious disease. However, the influence of this environment on host-pathogen interactions remains poorly understood. Here we report our results from the STL-IMMUNE study flown aboard Space Shuttle mission STS-131, which investigated multi-omic responses (transcriptomic, proteomic) of human intestinal epithelial cells to infection with Salmonella Typhimurium when both host and pathogen were simultaneously exposed to spaceflight. To our knowledge, this was the first in-flight infection and dual RNA-seq analysis using human cells.

Early development of fern gametophytes in microgravity.

Dormant spores of the fern Ceratopteris richardii were flown on Shuttle mission STS-93 to evaluate the effects of micro-g on their development and on their pattern of gene expression. Prior to flight the spores were sterilized and sown into one of two environments: (1) Microscope slides in a video-microscopy module; and (2) Petri dishes. All spores were then stored in darkness until use. Spore germination was initiated on orbit after exposure to light. For the spores on microscope slides, cell level changes were recorded through the clear spore coat of the spores by video microscopy. After their exposure to light, spores in petri dishes were frozen in orbit at four different time points during which on earth gravity fixes the polarity of their development. Spores were then stored frozen in Biological Research in Canister units until recovery on earth. The RNAs from these cells and from 1-g control cells were extracted and analyzed on earth after flight to assay changes in gene expression. Video microscopy results revealed that the germinated spores developed normally in microgravity, although the polarity of their development, which is guided by gravity on earth, was random in space. Differential Display-PCR analyses of RNA extracted from space-flown cells showed that there was about a 5% change in the pattern of gene expression between cells developing in micro-g compared to those developing on earth.

Continuous oxygen monitoring of mammalian cell growth on space shuttle mission STS-93 with a novel radioluminescent oxygen sensor.

A compact, flow-through oxygen sensor device based on luminescence quenching was used to monitor dissolved oxygen levels during mammalian cell growth on the STS-93 mission of the Columbia space shuttle. Excitation of an oxygen-sensitive ruthenium complex was provided by a radioluminescent light source (0.9 mm in diameter, 2.5 mm long), and the intensity of the resulting luminescence was measured by a simple photodiode detector. The use of radioluminescence for the excitation light source is a unique approach that provides many features important for long-term and remote monitoring applications. For the spaceflight experiment, human lung fibroblast cells (WI-38) were grown in hollow-fiber bioreactors. Oxygen concentration was measured in the flow path both before and after the bioreactor cartridge in order to gain information about the metabolism of the cells. The sensor was found to be nonperturbing to cell growth and withstood the challenging physical conditions of shuttle launch and landing while maintaining a stable calibration function. In addition, the sensor provided physically meaningful oxygen predictions.