Mechanobiologic Research in a Microgravity Environment Bioreactor

A current problem in tissue culturing technology is the unavailability of an effective Bioreactor for the in vitro cultivation of cells and explants. It has, in fact, proved extremely difficult to promote the high-density three-dimensional in vitro growth of human tissues that have been removed from the body and deprived of their normal in vivo vascular sources of nutrients and gas exchange. A variety of tissue explants can be maintained for a short period of time on a supportive collagen matrix surrounded by culture medium. But this system provides only limited mass transfer of nutrients and wastes through the tissue, and gravity-induced sedimentation prevents complete three- dimensional cell-cell and cell-matrix interactions. Several devices presently on the market have been used with only limited success since each has limitations, which restrict usefulness and versatility. Further, no Bioreactor or culture vessel is known that will allow for unimpeded growth of three dimensional cellular aggregates or tissue. Extensive research on the effect of mechanical stimuli on cell metabolism suggests that tissues may respond to mechanical stimulation via loading-induced flow of the interstitial fluids. During the culture, cells are subject to a flow of culture medium. Flow properties such as flow field, flow regime (e.g. turbulent or laminar), flow pattern (e.g. circular), entity and distribution of the shear stress acting on the cells greatly influence fundamental aspects of cell function, such as regulation and gene expression. This has been demonstrated for endothelial cells and significant research efforts are underway to elucidate these mechanisms in various other biological systems. Local fluid dynamics is also responsible of the mass transfer of nutrients and catabolites as well as oxygenation through the tissue. Most of the attempts to culture tissue-engineered constructs in vitro have utilized either stationary cultures or systems generating relatively small mechanical forces. For example, cartilage constructs have been cultured in spinner flasks under mixed or unmixed conditions, in simulated and in real microgravity. In these mixing studies, however, it is difficult to definitively quantify the effects of mixing-induced mechanical forces from those of convection-enhanced transport of nutrients to and of catabolites away from the cells. At the state of the art, the presence of a more controlled mechanical environment may be the condition required in order to study the biochemical and mechanical response of these biological systems. Such a controlled environment could lead to an advanced fluid dynamic design of the culture chamber that could both enhance the local mass transfer phenomena and match the needs of specific macroscopic mechanical effects in tissue development. The bioreactor is an excellent example of how the skills and resources of two distinctly different fields can complement each other. Microgravity can be used to enhance the formation of tissue like aggregates in specially designed bioreactors. Theoretical and experimental projects are under way to improve cell culture techniques using microgravity conditions experienced during space flights. Bioreactors usable under space flight conditions impose constructional principles which are different from those intended solely for ground applications. The Columbus Laboratory as part of the International Space Station (ISS) will be an evolving facility in low Earth orbit. Its mission is to support scientific, technological, and commercial activities in space. A goal of this research is to design a unique bioreactor for use sequentially from ground research to space research. One of the particularities of the simulated microgravity obtained through time averaging of the weight vector is that by varying the rotational velocity the same results can be obtained with a different value of g. One of the first applications of this technique in space biology was in fact the Rotating Wall Vessel developed by NASA, and originally designed to protect cell culture from the high shear forces generated during the launch and the landing of the Space Shuttle. A Bioreactor that is used both for ground and flight experiments provides the additional benefit of isolating dependent variable of gravity. This continuity will provide a means to compare results to a control experiment.