Human Tissue-Like Cellular Assemblies Grown for Respiratory Studies

One of the central objectives of this project was the development and characterization of a 3D mineralized tissue model system in which the effects of mechanical load (e.g., compression loading, tension, vibration, etc.) on the cellular responses of osteoblasts and osteoclasts could be investigated. After introducing mineralization agents to the culture, the constructs take on a bone-like appearance and have a more rigid structure suitable for being tested. Testing of the mineralized constructs confirmed the presence of calcium through a crystalline matrix histochemical stain. The central core is void of necrotic material, instead filled by a crystalline matrix with embedded nucleated cells. Remarkably, the nucleated cells do not express osteoblast markers, indicating differentiation to the in vivo cell type known as the osteocyte. In addition, as is characteristic to native periosteum, osteoclast precursor cells were imaged and proven to naturally arrange as an outer layer of the mineralized bone tissue construct. Development of this model will provide a unique venue for testing proposed countermeasures to space flight-induced bone loss. It will also allow a mechanistic approach in the modulation of cell signaling at the cellular level within the bone matrix. The Development And Characterization Of A Three-Dimensional Tissue Culture Model Of Bone is a technology readiness level (TRL) 6 (system/subsystem prototype demonstrated in a relevant environment). The innovation is now available for your company to license. Please note that NASA does not manufacture products itself for commercial sale.

The NASA developed Micro-Organ Device (MOD) platform technology is a small, lightweight, and reproducible in vitro drug screening model that can inexpensively biomimic different mammalian tissues for a multitude of applications. The technology is automated and imposes minimal demands for resources (power, analytes, and fluids). The MOD technology uses titanium tetra(isopropoxide) to bond a microscale support to a substrate and uses biopattering and 3D tissue bioprinting on a microfluidic microchip to eliminate variations in local seeding density while minimizing selection pressure. With the MOD, pharmaceutical companies can test more candidates and concentrate on those with more promise therefore, reducing R&D overall cost. This innovation overcomes major disadvantages of conventional in vitro and in vivo experimentation for purposes of investigating effects of medicines, toxins, and possibly other foreign substances. For example, the MOD platform technology could host life-like miniature assemblies of human cells and the effects observed in tests performed could potentially be extrapolated more readily to humans than could effects observed in conventional in vitro cell cultures, making it possible to reduce or eliminate experimentation on animals. The automated NASA developed technology with minimal footprint and power requirements, micro-volumes of fluids and waste, high throughput and parallel analyses on the same chip, will advance the research and development for new drugs and materials.

The apparatus comprises a randomized gravity vector multiphasic culture system with a self-feeding growth module, an optionally disposable nutrient module, and a removable AIMR chamber that delivers a pulsating multivariant field to the contents of the culture system. It produces overlapping or fluctuating alternating ionic magnetic resonance frequencies at one or more modal intervals ranging from about 7.8 Hz to about 59.9 Hz to the cell chamber. The apparatus may yield better regulation that can be manipulated to allow for increased rate of cell growth, faster differentiation, increased cell fidelity, and the induction or suppression of selective physiological genes involved in directing cellular differentiation and dedifferentiation. The use of an AIMR field may provide a significant improvement over existing bioreactors, including pulsating electromagnetic field (PEMF) and time-variance electromagnetic field (TVEMF) cellular growth induced systems, in that AIMR incorporates the modulation of cellular transcription. The AIMR system utilizes pre-sterilized disposable modules and a removable alternating ionic magnetic resonance chamber, reducing the hazard for contamination, allowing scientists to implement physiological and homeostatic parameters similar to a naturally occurring physiological system.

Current scaffold designs and materials do not provide all of the appropriate cues necessary to mimic in-vivo conditions for tissue engineering and stem cell engineering applications. It has been hypothesized that many biomaterials, such as bone, muscle, brain and heart tissue exhibit piezoelectric and ferroelectric properties. Typical cell seeding environments incorporate biochemical cues and more recently mechanical stimuli, however, electrical cues have just recently been incorporated in standard in-vitro examinations. In order to develop their potential further, novel scaffolds are required to provide adequate cues in the in-vitro environment to direct stem cells to differentiate down controlled pathways or develop novel tissue constructs. This invention is for a scaffold that provides for such cues by mimicking the native biological environment, including biochemical, topographical, mechanical and electrical cues.

The miniature bioreactor system was developed to provide the capabilities for NASA to perform cell studies in space and then provide results back to investigators on Earth with minimal tools and cost. The miniature bioreactor system has the potential to also be used on Earth as a laboratory bench-top cell culturing system without the need for expensive equipment and reagents. The system can be operated under computer control to reduce the operator handling and to reduce result variations. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere and temperature, and control subsystems. The system can be used to culture both anchorage dependent and suspension cells (prokaryotic or eukaryotic cell types). Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. The miniature bioreactor system for cell culturing has applications in pharmaceutical drug screening and cell culture studies.