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At least 73 records · Page 4

A model for the kinetics of homotypic cellular aggregation under static conditions

We present the formulation and testing of a mathematical model for the kinetics of homotypic cellular aggregation. The model considers cellular aggregation under no-flow conditions as a two-step process. Individual cells and cell aggregates 1) move on the tissue culture surface and 2) collide with other cells (or aggregates). These collisions lead to the formation of intercellular bonds. The aggregation kinetics are described by a system of coupled, nonlinear ordinary differential equations, and the collision frequency kernel is derived by extending Smoluchowski's colloidal flocculation theory to cell migration and aggregation on a two-dimensional surface. Our results indicate that aggregation rates strongly depend upon the motility of cells and cell aggregates, the frequency of cell-cell collisions, and the strength of intercellular bonds. Model predictions agree well with data from homotypic lymphocyte aggregation experiments using Jurkat cells activated by 33B6, an antibody to the beta 1 integrin. Since cell migration speeds and all the other model parameters can be independently measured, the aggregation model provides a quantitative methodology by which we can accurately evaluate the adhesivity and aggregation behavior of cells.

Non-NASA Center↗

Overview of the Life Science Glovebox (LSG) Facility and the Research Performed in the LSG

The Life Science Glovebox (LSG) is a rack facility currently under development with a projected availability for International Space Station (ISS) utilization in the FY2018 timeframe. Development of the LSG is being managed by the Marshal Space Flight Center (MSFC) with support from Ames Research Center (ARC) and Johnson Space Center (JSC). The MSFC will continue management of LSG operations, payload integration, and sustaining following delivery to the ISS. The LSG will accommodate life science and technology investigations in a "workbench" type environment. The facility has a.Ii enclosed working volume that is held at a negative pressure with respect to the crew living area. This allows the facility to provide two levels of containment for handling Biohazard Level II and lower biological materials. This containment approach protects the crew from possible hazardous operations that take place inside the LSG work volume. Research investigations operating inside the LSG are provided approximately 15 cubic feet of enclosed work space, 350 watts of28Vdc and l IOVac power (combined), video and data recording, and real time downlink. These capabilities will make the LSG a highly utilized facility on ISS. The LSG will be used for biological studies including rodent research and cell biology. The LSG facility is operated by the Payloads Operations Integration Center at MSFC. Payloads may also operate remotely from different telescience centers located in the United States and different countries. The Investigative Payload Integration Manager (IPIM) is the focal to assist organizations that have payloads operating in the LSG facility. NASA provides an LSG qualification unit for payload developers to verify that their hardware is operating properly before actual operation on the ISS. This poster will provide an overview of the LSG facility and a synopsis of the research that will be accomplished in the LSG. The authors would like to acknowledge Ames Research Center, Johnson Space Center, Teledyne Brown Engineering, MOOG-Bradford Engineering and the entire LSG Team for their inputs into this abstract.

Cole, J. Michael↗

Life Sciences Glovebox (LSG)

The Life Sciences Glovebox (LSG) accommodates life science and technology investigations in a “workbench” type environment. It is used for biological studies, including rodent research and cell biology. The facility has an enclosed Work Volume (WV) that is held at a negative pressure with respect to the crew living area. This allows the facility to provide two levels of containment for handling Biohazard Level II and lower biological materials. The LSG facility is operated by the Payloads Operations Integration Center at MSFC. Payloads may also operate remotely from different telescience centers located in the United States and different countries. The Glovebox Integration Engineer (GIE) is the focal to assist organizations that have payloads operating in the LSG facility. NASA provides an LSG engineering unit for payload developers, fit checks, and to verify that the hardware is operating properly before beginning actual operations on the ISS.

biology↗

Fluorescence activated cell sorting.

An instrument has been developed for sorting biological cells. The cells are rendered differentially fluorescent and incorporated into a small liquid stream illuminated by a laser beam. The cells pass sequentially through the beam, and fluorescent light from the cells gives rise to electrical signals. The stream is broken into a series of uniform size drops downstream of the laser. The cell signals are used to give appropriate electrostatic charges to drops containing the cells. The drops then pass between two charged plates and are deflected to appropriate containers. The system has proved capable of providing fractions containing large numbers of viable cells highly enriched in a particular functional type.

Bonner, W. A.↗

Cell Partition in Two Polymer Aqueous Phases

Partition of biological cells in two phase aqueous polymer systems is recognized as a powerful separation technique which is limited by gravity. The synthesis of new, selective polymer ligand conjugates to be used in affinity partition separations is of interest. The two most commonly used polymers in two phase partitioning are dextran and polyethylene glycol. A thorough review of the chemistry of these polymers was begun, particularly in the area of protein attachment. Preliminary studies indicate the importance in affinity partitioning of minimizing gravity induced randomizing forces in the phase separation process. The PEG-protein conjugates that were prepared appear to be ideally suited for achieving high quality purifications in a microgravity environment. An interesting spin-off of this synthetic work was the observation of catalytic activity for certain of our polymer derivatives.

Harris, J. M.↗

Cell and Molecular Biology of Ataxia Telangiectasia Heterozygous Human Mammary Epithelial Cells Irradiated in Culture

Autologous isolates of cell types from obligate heterozygotes with the autosomal disorder ataxia-telangiectasia (A-T)were used to begin a tissue culture model for assessing pathways of radiation-induced cancer formation in this target tissue. This was done by establishing cultures of stromal fibroblasts and long-term growth human mammary epithelial cells (HMEC) in standard 2-dimensional tissue culture in order to establish expression of markers detailing early steps of carcinogenesis. The presumptive breast cancer susceptibility of A-T heterozygotes as a sequel to damage caused by ionizing radiation provided reason to study expression of markers in irradiated HMEC. Findings from our study with HMEC have included determination of differences in specific protein expression amongst growth phase (e.g., log vs stationary) and growth progression (e.g., pass 7 vs pass 9), as well as differences in morphologic markers within populations of irradiated HMEC (e.g., development of multinucleated cells).

Richmond, Robert C.↗

Propulsion of synthetic protocells and coacervates driven by biochemical catalysis

An important area in the design of biomolecular materials is the mimicry of cellular function through the assembly of molecular components into synthetic cells, or protocells. Through compartmentalization and subsequent evolution, biological cells have achieved the goals of specificity, amplification, and response. By assembling cell-like structures that mimic the organization and size of a natural cell but that incorporate novel components and respond to synthetic triggers, we can extend their capabilities beyond what is currently achievable in biology. For applications in energy, these include the ability to convert chemical to mechanical energy, and to enable the response of sensors and responders once a threshold has been reached. This technical report summarizes progress made on this project as of Dec 31, 2023.

36 MATERIALS SCIENCE↗

New polymers for low-gravity purification of cells by phase partitioning

A potentially powerful technique for separating different biological cell types is based on the partitioning of these cells between the immiscible aqueous phases formed by solution of certain polymers in water. This process is gravity-limited because cells sediment rather than associate with the phase most favored on the basis of cell-phase interactions. In the present contract we have been involved in the synthesis of new polymers both to aid in understanding the partitioning process and to improve the quality of separations. The prime driving force behind the design of these polymers is to produce materials which will aid in space experiments to separate important cell types and to study the partitioning process in the absence of gravity (i.e., in an equilibrium state).

Harris, J. M.↗

Bone Biomarkers on the Pathway to Effective Spaceflight Countermeasures

Osteocyte cells are the most abundant yet least understood bone cell type in the human body. However, recent discovers in osteocyte cell biology have shed light on their importance as key mechanosensing cells regulating the bone remodeling process. Thus, we propose the first ever in vitro gene expression evaluation of osteocytes exposed to simulated microgravity to determine mechanistic pathways of their gravity sensing ability. Improved understanding of the fundamental mechanisms at the osteocyte cellular level may lead to improved treatment options to mitigate the effects of bone loss encountered by astronauts on long duration space missions and provide tailored treatment options for maintaining bone strength of immobilized/partially paralyzed patients here on Earth. Aim 1: Characterize the gene expression patterns and protein levels following exposure of murine osteocytelike cell line (MLO-Y4) to simulated microgravity using the NASA Rotating Wall Vessel (RWV) Bioreactor. Osteocytes are theorized to be the mechanosensors and transducers of mechanical load for bones, yet the biological mechanism of this action remains elusive. We propose to investigate the genetic regulation of the mechanism of the MLO-Y4 cell in the NASA Bioreactor as it is the accepted ground-based analog for simulating vector averaged microgravity.

Spatz, Jordan↗

Electrophoresis technology experiment MA-011

Experiment MA-011, electrophoresis technology, was designed to test electrophoresis hardware that would continue the development of technology for electrophoretic separation of materials in the near zero g environment of space. The experimental hardware generally functioned as planned. Frozen live cells were successfully transported into space, electrophoretic processing was performed, and viable cells were returned to earth. A separation of the three types of fixed red blood cells (rabbit, human, and horse) was demonstrated. The human lymphocytes, however, showed no apparent migration. The separation of human kidney cells produced the most exciting data. Analysis shows electrophoretic separation throughout the entire column with at least four bands of viable cells. The isotachophoresis experiment definitely demonstrated the isotachophoretic separation of biological cells in a near zero g environment.

Allen, R. E.↗

The August Krogh principle applies to plants

The Krogh principle refers to the use of a large number of animals to study the large number of physiological problems, rather than limiting study to a particular organism for all problems. There may be organisms that are more suited to study of a particular problem than others. This same principle applies to plants. The authors are concerned with the recent trend in plant biology of using Arabidopsis thaliana as the "organism of choice." Arabidopsis is an excellent organism for molecular genetic research, but other plants are superior models for other research areas of plant biology. The authors present examples of the successful use of the Krogh principle in plant cell biology research, emphasizing the particular characteristics of the selected research organisms that make them the appropriate choice.

NASA Discipline Plant Biology↗

ARC Cell Science Validation (CS-V) Payload Overview

Automated cell biology system for laboratory and International Space Station (ISS) National Laboratory research. Enhanced cell culture platform that provides undisturbed culture maintenance, including feedback temperature control, medical grade gas supply, perfusion nutrient delivery and removal of waste, and automated experiment manipulations. Programmable manipulations include: media feeds change out, injections, fraction collections, fixation, flow rate, and temperature modification within a one-piece sterile barrier flow path. Cassette provides 3 levels of containment and allows Crew access to the bioculture chamber and flow path assembly for experiment initiation, refurbishment, or sample retrieval and preservation.

Space Biology↗

Cameleon calcium indicator reports cytoplasmic calcium dynamics in Arabidopsis guard cells

Cytoplasmic free calcium ([Ca2+]cyt) acts as a stimulus-induced second messenger in plant cells and multiple signal transduction pathways regulate [Ca2+]cyt in stomatal guard cells. Measuring [Ca2+]cyt in guard cells has previously required loading of calcium-sensitive dyes using invasive and technically difficult micro-injection techniques. To circumvent these problems, we have constitutively expressed the pH-independent, green fluorescent protein-based calcium indicator yellow cameleon 2.1 in Arabidopsis thaliana (Miyawaki et al. 1999; Proc. Natl. Acad. Sci. USA 96, 2135-2140). This yellow cameleon calcium indicator was expressed in guard cells and accumulated predominantly in the cytoplasm. Fluorescence ratio imaging of yellow cameleon 2.1 allowed time-dependent measurements of [Ca2+]cyt in Arabidopsis guard cells. Application of extracellular calcium or the hormone abscisic acid (ABA) induced repetitive [Ca2+]cyt transients in guard cells. [Ca2+]cyt changes could be semi-quantitatively determined following correction of the calibration procedure for chloroplast autofluorescence. Extracellular calcium induced repetitive [Ca2+]cyt transients with peak values of up to approximately 1.5 microM, whereas ABA-induced [Ca2+]cyt transients had peak values up to approximately 0.6 microM. These values are similar to stimulus-induced [Ca2+]cyt changes previously reported in plant cells using ratiometric dyes or aequorin. In some guard cells perfused with low extracellular KCl concentrations, spontaneous calcium transients were observed. As yellow cameleon 2.1 was expressed in all guard cells, [Ca2+]cyt was measured independently in the two guard cells of single stomates for the first time. ABA-induced, calcium-induced or spontaneous [Ca2+]cyt increases were not necessarily synchronized in the two guard cells. Overall, these data demonstrate that that GFP-based cameleon calcium indicators are suitable to measure [Ca2+]cyt changes in guard cells and enable the pattern of [Ca2+]cyt dynamics to be measured with a high level of reproducibility in Arabidopsis cells. This technical advance in combination with cell biological and molecular genetic approaches will become an invaluable tool in the dissection of plant cell signal transduction pathways.

Non-NASA Center↗

Perfusion Bioreactor Module

Perfusion bioreactor module, self-contained, closed-loop cell-culture system that operates in microgravity or on Earth. Equipment supports growth or long-term maintenance of cultures of human or other fragile cells for experiments in basic cell biology or process technology. Designed to support proliferation (initially at exponential rates of growth) of cells in complex growth medium and to maintain confluent cells in defined medium under conditions optimized to permit or encourage selected functions of cells, including secretion of products of cells into medium.

Morrison, Dennis R.↗

Multistage Magnetic Separator of Cells and Proteins

The multistage electromagnetic separator for purifying cells and magnetic particles (MAGSEP) is a laboratory apparatus for separating and/or purifying particles (especially biological cells) on the basis of their magnetic susceptibility and magnetophoretic mobility. Whereas a typical prior apparatus based on similar principles offers only a single stage of separation, the MAGSEP, as its full name indicates, offers multiple stages of separation; this makes it possible to refine a sample population of particles to a higher level of purity or to categorize multiple portions of the sample on the basis of magnetic susceptibility and/or magnetophoretic mobility. The MAGSEP includes a processing unit and an electronic unit coupled to a personal computer. The processing unit includes upper and lower plates, a plate-rotation system, an electromagnet, an electromagnet-translation system, and a capture-magnet assembly. The plates are bolted together through a roller bearing that allows the plates to rotate with respect to each other. An interface between the plates acts as a seal for separating fluids. A lower cuvette can be aligned with as many as 15 upper cuvette stations for fraction collection during processing. A two-phase stepping motor drives the rotation system, causing the upper plate to rotate for the collection of each fraction of the sample material. The electromagnet generates a magnetic field across the lower cuvette, while the translation system translates the electromagnet upward along the lower cuvette. The current supplied to the electromagnet, and thus the magnetic flux density at the pole face of the electromagnet, can be set at a programmed value between 0 and 1,400 gauss (0.14 T). The rate of translation can be programmed between 5 and 2,000 m/s so as to align all sample particles in the same position in the cuvette. The capture magnet can be a permanent magnet. It is mounted on an arm connected to a stepping motor. The stepping motor rotates the arm to position the capture magnet above the upper cuvette into which a fraction of the sample is collected. The electronic unit includes a power switch, power-supply circuitry that accepts 110-Vac input power, an RS-232 interface, and status lights. The personal computer runs the MAGSEP software and controls the operation of the MAGSEP through the RS-232 interface. The status of the power, the translating electromagnet, the capture magnet, and the rotation of the upper plate are indicated in a graphical user interface on the computer screen.

Barton, Ken↗

Use of an adaptable cell culture kit for performing lymphocyte and monocyte cell cultures in microgravity

The results of experiments performed in recent years on board facilities such as the Space Shuttle/Spacelab have demonstrated that many cell systems, ranging from simple bacteria to mammalian cells, are sensitive to the microgravity environment, suggesting gravity affects fundamental cellular processes. However, performing well-controlled experiments aboard spacecraft offers unique challenges to the cell biologist. Although systems such as the European 'Biorack' provide generic experiment facilities including an incubator, on-board 1-g reference centrifuge, and contained area for manipulations, the experimenter must still establish a system for performing cell culture experiments that is compatible with the constraints of spaceflight. Two different cell culture kits developed by the French Space Agency, CNES, were recently used to perform a series of experiments during four flights of the 'Biorack' facility aboard the Space Shuttle. The first unit, Generic Cell Activation Kit 1 (GCAK-1), contains six separate culture units per cassette, each consisting of a culture chamber, activator chamber, filtration system (permitting separation of cells from supernatant in-flight), injection port, and supernatant collection chamber. The second unit (GCAK-2) also contains six separate culture units, including a culture, activator, and fixation chambers. Both hardware units permit relatively complex cell culture manipulations without extensive use of spacecraft resources (crew time, volume, mass, power), or the need for excessive safety measures. Possible operations include stimulation of cultures with activators, separation of cells from supernatant, fixation/lysis, manipulation of radiolabelled reagents, and medium exchange. Investigations performed aboard the Space Shuttle in six different experiments used Jurkat, purified T-cells or U937 cells, the results of which are reported separately. We report here the behaviour of Jurkat and U937 cells in the GCAK hardware in ground-based investigations simulating the conditions expected in the flight experiment. Several parameters including cell concentration, time between cell loading and activation, and storage temperature on cell survival were examined to characterise cell response and optimise the experiments to be flown aboard the Space Shuttle. Results indicate that the objectives of the experiments could be met with delays up to 5 days between cell loading into the hardware and initial in flight experiment activation, without the need for medium exchange. Experiment hardware of this kind, which is adaptable to a wide range of cell types and can be easily interfaced to different spacecraft facilities, offers the possibility for a wide range of experimenters successfully and easily to utilise future flight opportunities.

short duration↗

Kingdoms in turmoil

How should the world's living organisms be classified? Into how many kingdoms should they be grouped? Scientists have been grappling with these questions since the time of Aristotle, drawing on a broad base of biological characteristics for clues. The fossil record, visible traits of living organisms and, more recently, results from cell biology have all shaped theories of biological classification. But last year a new and controversial concept emerged: a classification of life based solely on molecular traits. The focal point of the controversy is a tree of life, or "phylogeny", devised by Carl Woese of the University of Illinois, Otto Kandler of the University of Munich and Mark Wheelis of the University of California. The tree is unusual because, unlike all previous schemes, it is constructed solely from biochemical data such as DNA sequences rather than a range of different organism characteristics. But that is not all. The scheme also challenges the idea that life on Earth is best divided into five kingdoms, with the main split being between bacteria and all other organisms. Woese and his colleagues create three main groupings by dividing the bacteria in two and unifying all other organisms.

NASA Program Exobiology↗