Engineering Papers⌕ Search

SEARCH · Engineering Papers

Results for “cells”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 847 records · Page 47
All sourcesNASA NTRSDOE OSTI
NASA NTRS2004

Utility of Thin-Film Solar Cells on Flexible Substrates for Space Power

The thin-film solar cell program at NASA GRC is developing solar cell technologies for space applications which address two critical metrics: specific power (power per unit mass) and launch stowed volume. To be competitive for many space applications, an array using thin film solar cells must significantly increase specific power while reducing stowed volume when compared to the present baseline technology utilizing crystalline solar cells. The NASA GRC program is developing two approaches. Since the vast majority of the mass of a thin film solar cell is in the substrate, a thin film solar cell on a very lightweight flexible substrate (polymer or metal films) is being developed as the first approach. The second approach is the development of multijunction thin film solar cells. Total cell efficiency can be increased by stacking multiple cells having bandgaps tuned to convert the spectrum passing through the upper cells to the lower cells. Once developed, the two approaches will be merged to yield a multijunction, thin film solar cell on a very lightweight, flexible substrate. The ultimate utility of such solar cells in space require the development of monolithic interconnections, lightweight array structures, and ultra-lightweight support and deployment techniques.

Dickman, J. E.↗
NASA NTRS2003

Retinal ganglion cell projections to the hamster suprachiasmatic nucleus, intergeniculate leaflet, and visual midbrain: bifurcation and melanopsin immunoreactivity

The circadian clock in the suprachiasmatic nucleus (SCN) receives direct retinal input via the retinohypothalamic tract (RHT), and the retinal ganglion cells contributing to this projection may be specialized with respect to direct regulation of the circadian clock. However, some ganglion cells forming the RHT bifurcate, sending axon collaterals to the intergeniculate leaflet (IGL) through which light has secondary access to the circadian clock. The present studies provide a more extensive examination of ganglion cell bifurcation and evaluate whether ganglion cells projecting to several subcortical visual nuclei contain melanopsin, a putative ganglion cell photopigment. The results showed that retinal ganglion cells projecting to the SCN send collaterals to the IGL, olivary pretectal nucleus, and superior colliculus, among other places. Melanopsin-immunoreactive (IR) ganglion cells are present in the hamster retina, and some of these cells project to the SCN, IGL, olivary pretectal nucleus, or superior colliculus. Triple-label analysis showed that melanopsin-IR cells bifurcate and project bilaterally to each SCN, but not to the other visual nuclei evaluated. The melanopsin-IR cells have photoreceptive characteristics optimal for circadian rhythm regulation. However, the presence of moderately widespread bifurcation among ganglion cells projecting to the SCN, and projection by melanopsin-IR cells to locations distinct from the SCN and without known rhythm function, suggest that this ganglion cell type is generalized, rather than specialized, with respect to the conveyance of photic information to the brain. Copyright 2003 Wiley-Liss, Inc.

NASA Discipline Neuroscience↗
NASA NTRS2005

Nonhumidified High-Temperature Membranes Developed for Proton Exchange Membrane Fuel Cells

Fuel cells are being considered for a wide variety of aerospace applications. One of the most versatile types of fuel cells is the proton-exchange-membrane (PEM) fuel cell. PEM fuel cells can be easily scaled to meet the power and space requirements of a specific application. For example, small 100-W PEM fuel cells are being considered for personal power for extravehicular activity suit applications, whereas larger PEM fuel cells are being designed for primary power in airplanes and in uninhabited air vehicles. Typically, PEM fuel cells operate at temperatures up to 80 C. To increase the efficiency and power density of the fuel cell system, researchers are pursuing methods to extend the operating temperature of the PEM fuel cell to 180 C. The most widely used membranes in PEM fuel cells are Nafion 112 and Nafion 117--sulfonated perfluorinated polyethers that were developed by DuPont. In addition to their relatively high cost, the properties of these membranes limit their use in a PEM fuel cell to around 80 C. The proton conductivity of Nafion membranes significantly decreases above 80 C because the membrane dehydrates. The useful operating range of Nafion-based PEM fuel cells can be extended to over 100 C if ancillary equipment, such as compressors and humidifiers, is added to maintain moisture levels within the membrane. However, the addition of these components reduces the power density and increases the complexity of the fuel cell system.

Kinder, James D.↗
NASA NTRS2006

Lithium-Ion Cell Charge Control Unit

Life-test data of Lithium-Ion battery cells is critical in order to establish their performance capabilities for NASA missions and Exploration goals. Lithium-ion cells have the potential to replace rechargeable alkaline cells in aerospace applications, but they require a more complex charging scheme than is typically required for alkaline cells. To address these requirements in our Lithium-Ion Cell Test Verification Program, a Lithium-Ion Cell Charge Control Unit was developed by NASA Glenn Research Center (GRC). This unit gives researchers the ability to test cells together as a pack, while allowing each cell to charge individually. This allows the inherent cell-to-cell variations to be addressed on a series string of cells and results in a substantial reduction in test costs as compared to individual cell testing. The Naval Surface Warfare Center at Crane, Indiana developed a power reduction scheme that works in conjunction with the Lithium-Ion Cell Charge Control Unit. This scheme minimizes the power dissipation required by the circuitry to prolong circuit life and improve its reliability.

Reid, Concha↗
NASA NTRS2010

Oscillating Cell Culture Bioreactor

To better exploit the principles of gas transport and mass transport during the processes of cell seeding of 3D scaffolds and in vitro culture of 3D tissue engineered constructs, the oscillatory cell culture bioreactor provides a flow of cell suspensions and culture media directly through a porous 3D scaffold (during cell seeding) and a 3D construct (during subsequent cultivation) within a highly gas-permeable closed-loop tube. This design is simple, modular, and flexible, and its component parts are easy to assemble and operate, and are inexpensive. Chamber volume can be very low, but can be easily scaled up. This innovation is well suited to work with different biological specimens, particularly with cells having high oxygen requirements and/or shear sensitivity, and different scaffold structures and dimensions. The closed-loop changer is highly gas permeable to allow efficient gas exchange during the cell seeding/culturing process. A porous scaffold, which may be seeded with cells, is fixed by means of a scaffold holder to the chamber wall with scaffold/construct orientation with respect to the chamber determined by the geometry of the scaffold holder. A fluid, with/without biological specimens, is added to the chamber such that all, or most, of the air is displaced (i.e., with or without an enclosed air bubble). Motion is applied to the chamber within a controlled environment (e.g., oscillatory motion within a humidified 37 C incubator). Movement of the chamber induces relative motion of the scaffold/construct with respect to the fluid. In case the fluid is a cell suspension, cells will come into contact with the scaffold and eventually adhere to it. Alternatively, cells can be seeded on scaffolds by gel entrapment prior to bioreactor cultivation. Subsequently, the oscillatory cell culture bioreactor will provide efficient gas exchange (i.e., of oxygen and carbon dioxide, as required for viability of metabolically active cells) and controlled levels of fluid dynamic shear (i.e., as required for viability of shear-sensitive cells) to the developing engineered tissue construct. This bioreactor was recently utilized to show independent and interactive effects of a growth factor (IGF-I) and slow bidirectional perfusion on the survival, differentiation, and contractile performance of 3D tissue engineering cardiac constructs. The main application of this system is within the tissue engineering industry. The ideal final application is within the automated mass production of tissue- engineered constructs. Target industries could be both life sciences companies as well as bioreactor device producing companies.

Freed, Lisa E.↗
NASA NTRS2010

Performance Characterization of High Energy Commercial Lithium-ion Cells

The NASA Glenn Research Center Electrochemistry Branch performed characterization of commercial lithium-ion cells to determine the cells' performance against Exploration Technology Development Program (ETDP) Key Performance Parameters (KPP). The goals of the ETDP Energy Storage Project require significant improvements in the specific energy of lithium-ion technology over the state-of-the-art. This work supports the high energy cell development for the Constellation customer Lunar Surface Systems (LSS). In support of these goals, testing was initiated in September 2009 with high energy cylindrical cells obtained from Panasonic and E-One Moli. Both manufacturers indicated the capability of their cells to deliver specific energy of at least 180 Wh/kg or higher. Testing is being performed at the NASA Glenn Research Center to evaluate the performance of these cells under temperature, rate, and cycling conditions relevant to the ETDP goals for high energy cells. The cell-level specific energy goal for high energy technology is 180 Wh/kg at a C/10 rate and 0 C. The threshold value is 165 Wh/kg. The goal is to operate for at least 2000 cycles at 100 percent DOD with greater than 80 percent capacity retention. The Panasonic NCR18650 cells were able to deliver nearly 200 Wh/kg at the aforementioned conditions. The E-One Moli ICR18650J cells also met the specific energy goal by delivering 183 Wh/kg. Though both cells met the goal for specific energy, this testing was only one portion of the testing required to determine the suitability of commercial cells for the ETDP. The cells must also meet goals for cycle life and safety. The results of this characterization are summarized in this report.

Schneidegger, Brianne T.↗
NASA NTRS2001

Whole Blood Activation Results in Enhanced Detection of T Cell and Monocyte Cytokine Production by Flow Cytometry

An excellent monitor of the immune balance of peripheral circulating cells is to determine their cytokine production patterns in response to stimuli. Using flow cytometry a positive identification of cytokine producing cells in a mixed culture may be achieved. Recently, the ability to assess cytokine production following a wholeblood activation culture has been described. We compared whole blood culture to standard PBMC culture and determined the individual cytokine secretion patterns for both T cells and monocytes via flow cytometry. For T cells cytokine assessment following PMA +ionomycin activation: (1) a significantly greater percentages of T cells producing IFNgamma and IL-2 were observed following whole-blood culture; (2) altered T cell cytokine production kinetics were observed by varying whole blood culture times. In addition, a four-color cytometric analysis was used to allow accurate phenotyping and quantitation of cytokine producing lymphocyte populations. Using this technique we found IFNgamma production to be significantly elevated in the CD3+/CD8+ T cell population as compared to the CD3+/CD8- population following five hours of whole blood activation. Conversely, IL-2 and IL-10 production were significantly elevated in the CD3+/CD8- T cell population as compared to the CD3+/CD8+ population. Monocyte cytokine production was assessed in both culture systems following LPS activation for 24 hours. A three-color flow cytometric was used to assess two cytokines in conjunction with CD 14. The cytokine pairs used for analysis were IL-1a/IL-12, and IL-10ITNFa. Nearly all monocytes were stimulated to produce IL-1a, IL-12 and TNFalpha equally well in both culture systems. Monocyte production of IL-10 was significantly elevated following whole blood culture as compared to PBMC culture. IL-12 producing monocytes appeared to be a distinct subpopulation of the IL-1a producing set, whereas IL-10 and TNFa producing monocytes were largely mutually exclusive. IL-10 and TNFa producing monocytes may represent functionally different monocyte subsets with distinct functions. Whole blood culture eliminates the need to purify cell populations prior to culture and may have significant utility for the routine monitoring of the cytokine balances of the peripheral blood T cell and monocyte populations. In addition, there are distinct advantages to performing whole-blood (WB) activation as compared to PBMC activation. These advantages would include retaining all various cell-cell interactions as well as any soluble factors present in serum that influence cell activation. It is likely that the altered cytokine production observed following whole blood culture more accurately represents the in-vivo immune balance.

Sams, Clarence F.↗
NASA NTRSDate not supplied

CDKN1A/P21’S Role in Osteoprogenitor Fate Regulation in Response to Mechanical Loading: A Single Cell Resolution Investigation

Forces generated by gravity in load-bearing tissues such as bone marrow promote stem cell-based tissue-regenerative processes by increasing proliferation and differentiation of tissue progenitors. Conversely, in microgravity, mouse bone marrow mesenchymal and hematopoietic precursors down-regulate differentiation markers and up-regulate stemness maintenance genes. During microgravity, these transcriptomic changes are associated with Cdkn1a overexpression in osteoprogenitors, and further the homozygous deletion of the CDKN1a gene in the mouse model results in elevated stem-cell based regeneration of severed digits and ear-hole punches. In this work we sought to test the hypothesis that gravity mechanotransduction regulates mesenchymal stem cell derived osteoprogenitor based osteogenesis by modulating proliferation and differentiation fates at specific cell cycle stages via a p21/CDKN1a-regulated mechanism.. To test our hypothesis, we isolated primary mouse bone marrow osteoprogenitors from Cdkn1a-/- (null) or wildtype mice, cultured the cells for 7-days unloaded in osteogenic differentiation media, to allow commitment of the adherent cell population, and subsequently, cultured an additional 48-hours under cyclic stretch or static control loading conditions. After 48-hours of dynamic mechanical loading, we used a 10X Genomics Chromium/Single Cell controller to generate bar-coded single cell Illumina libraries and sequenced expressomes for 6,000 static control and 6,000 cyclic stretch cells for each genotype. Stage-specific single cell analyses show Cdkn1a-/- osteogenic cultures are more differentiated with fewer progenitors, and that cyclic stretch further promotes the conversion of progenitors to osteoblasts in both wildtype and Cdkn1a-/- backgrounds. The proportion of late mineralizing osteoblasts doubled in Cdkn1a-/- cultures, but is not affected by load. Finally, single cell expression of Cdkn1a, in the wildtype cells, is most strongly suppressed by cyclic stretch both in early and late osteoblasts, and minimally in the progenitor population. Collectively, our results support the hypothesis that Cdkn1a constitutively plays a mechano-reversible anti-proliferative role during stem cell-based bone tissue regeneration, and suggests a new molecular target to counter regenerative deficits caused by disuse.

Eduardo Almeida↗
NASA NTRSDate not supplied

Enabling Single Cell Research in Hollow Hydrogel Microparticles

Studying biology at the single cell level enables novel discoveries through ultra-high throughput identification and isolation of rare cells and interesting subpopulations. Encapsulation of single cells in hollow hydrogel microparticles, or PicoShells, enables easy visualization, manipulation, and assaying of single cells. The PicoShell’s porous hydrogel shell allows diffusion of nutrients and wastes into and out of the hollow liquid core, allowing uninhibited cell growth. The porous shell also allows diffusion of assay reagents, enabling solution changes, and multi-step assays. Finally, single cells and their progeny can be tracked through time, for example to measure single cell growth rates. However, PicoShells are fabricated using microfluidic techniques, which require specialized expertise, equipment, and facilities. This limits PicoShell’s widespread utility for single cell biology. For Saccharomyces cerevisiae and other desiccation-tolerant microbes, desiccating PicoShells containing cells of interest offers a solution, enabling those without microfluidic capabilities to obtain dried, shelf-stable PicoShells containing their cells of interest from a specialized microfluidics facility. Researchers could then rehydrate the PicoShells and perform their single cell assays, bypassing the microfluidic steps. Desiccated PicoShells could also be flown in space, where desiccation is already a common practice for microbes (i.e. BioSentinel and BioNutrients), bringing single cell analysis capabilities to space. In this work, we develop a method to desiccate PicoShells containing S. cerevisiae while maintaining characteristics similar to fresh, non-desiccated PicoShells. We show good microparticle morphology and hydrogel pore size as well as high yeast viability in line with previous studies. We also demonstrate sorting of a fast-growing yeast population using size-based filtration, as S. cerevisiae in PicoShells are able to physically stretch the microparticle as they grow, increasing the PicoShell diameter. These efforts contribute to the feasibility of leveraging microparticle PicoShells for single cell space biology research on Earth and in space.

Simon Ng↗
DOE OSTI2023

Establishing metrics to quantify spatial similarity in spherical and red blood cell distributions

As computational power increases and systems with millions of red blood cells can be simulated, it is important to note that varying spatial distributions of cells may affect simulation outcomes. Since a single simulation may not represent the ensemble behavior, many different configurations may need to be sampled to adequately assess the entire collection of potential cell arrangements. In order to determine both the number of distributions needed and which ones to run, we must first establish methods to identify well-generated, randomly placed cell distributions and to quantify distinct cell configurations. We utilize metrics to assess (1) the presence of any underlying structure to the initial cell distribution and (2) similarity between cell configurations. We propose the use of the radial distribution function to identify long-range structure in a cell configuration and apply it to a randomly distributed and structured set of red blood cells. To quantify spatial similarity between two configurations, we make use of the Jaccard index, and characterize sets of red blood cell and sphere initializations. As an extension to our work submitted to the International Conference on Computational Science, we significantly increase our data set size from 72 to 1048 cells, include a similar set of studies using spheres, compare the effects of varying sphere size, and utilize the Jaccard index distribution to probe sets of extremely similar configurations. Our results show that the radial distribution function can be used as a metric to determine long-range structure in both distributions of spheres and RBCs. We determine that the ideal case of spheres within a cube versus bi-concave shaped cells within a cylinder affects the shape of the Jaccard index distributions, as well as the range of Jaccard values, showing that both the shape of particle and the domain may play a role. Furthermore, we also find that the distribution is able to capture very similar configurations through Jaccard index values greater than 95% when appending several nearly identical configurations into the data set.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Tunicamycin-induced endoplasmic reticulum stress inhibits chemoresistance of FaDu hypopharyngeal carcinoma cells in 3D collagen I cultures and in vivo

Highlights: • 3D collagen I culture induces chemoresistance of FaDu hypopharyngeal carcinoma cells. • Tunicamycin-induced ER stress inhibits chemoresistance in collagen I cultures. • Tunicamycin-induced ER stress potentiates anticancer efficacy in vivo. • Tunicamycin-induced ER stress inhibits TGF-β1 and modifies integrin β1 glycosylation. The prognosis in patients with advanced head and neck squamous cell carcinoma (HNSCC) is widely affected by the resistance to chemotherapy. As a culture scaffold, collagen I was showed to promote CSC (cancer stem cell) properties of cancer cells which could be used as in vitro models to study the chemoresistance in HNSCC. Endoplasmic reticulum (ER) stress is a cellular stress condition which could affect tumor progression and promote the anti-tumor effects of certain drugs. However, the impact of ER stress on collagen I induced CSC properties and chemoresistance of HNSCC cells has not been addressed. In this study we investigated the effects of tunicamycin (TM) induced ER stress on the stemness and sensitivity to chemotherapeutic drugs of FaDu hypopharyngeal carcinoma cells in 3D (three-dimensional) collagen I cultures and mouse xenograft models. Our study revealed that Collagen I scaffold promoted CSC properties and increased G1 population of FaDu cells in 3D cultures, accompanied by maturation of integrin β1 and enhanced activated TGF-β1 concentration. Compared to 2D (two-dimensional) cultured cells, cells in 3D Collagen I scaffold exhibited significantly increased resistance to chemotherapeutic drugs of cisplatin and paclitaxel. Further analysis revealed that TM induced ER stress preferentially attenuated chemoresistance of FaDu cells in 3D collagen I, downregulated their CSC properties and TGF-β1 concentration and resulted in deglycosylation of integrin β1. TM was further evaluated in the mouse xenograft models and showed significant tumor growth inhibition in combination with paclitaxel than either TM or paclitaxel alone. Taken together, Our findings suggest that TM-induced ER stress potentiates anticancer efficacy of FaDu cells in 3D cultures and in vivo, and highlight implications for targeting chemotherapy-resistant cancer stem cells under ER stress conditions.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2023

Engineered molecular sensors for quantifying cell surface crowding

Cells mediate interactions with the extracellular environment through a crowded assembly of transmembrane proteins, glycoproteins and glycolipids on their plasma membrane. The extent to which surface crowding modulates the biophysical interactions of ligands, receptors, and other macromolecules is poorly understood due to the lack of methods to quantify surface crowding on native cell membranes. In this work, we demonstrate that physical crowding on reconstituted membranes and live cell surfaces attenuates the effective binding affinity of macromolecules such as IgG antibodies in a surface crowding-dependent manner. We combine experiment and simulation to design a crowding sensor based on this principle that provides a quantitative readout of cell surface crowding. Our measurements reveal that surface crowding decreases IgG antibody binding by 2 to 20 fold in live cells compared to a bare membrane surface. Our sensors show that sialic acid, a negatively charged monosaccharide, contributes disproportionately to red blood cell surface crowding via electrostatic repulsion, despite occupying only ~1% of the total cell membrane by mass. We also observe significant differences in surface crowding for different cell types and find that expression of single oncogenes can both increase and decrease crowding, suggesting that surface crowding may be an indicator of both cell type and state. Our high-throughput, single-cell measurement of cell surface crowding may be combined with functional assays to enable further biophysical dissection of the cell surfaceome.

60 APPLIED LIFE SCIENCES↗
NASA NTRS2002

Dynamics of cell proliferation in the adult dentate gyrus of two inbred strains of mice

The output potential of proliferating populations in either the developing or the adult nervous system is critically dependent on the length of the cell cycle (T(c)) and the size of the proliferating population. We developed a new approach for analyzing the cell cycle, the 'Saturate and Survive Method' (SSM), that also reveals the dynamic behaviors in the proliferative population and estimates of the size of the proliferating population. We used this method to analyze the proliferating population of the adult dentate gyrus in 60 day old mice of two inbred strains, C57BL/6J and BALB/cByJ. The results show that the number of cells labeled by exposure to BUdR changes dramatically with time as a function of the number of proliferating cells in the population, the length of the S-phase, cell division, the length of the cell cycle, dilution of the S-phase label, and cell death. The major difference between C57BL/6J and BALB/cByJ mice is the size of the proliferating population, which differs by a factor of two; the lengths of the cell cycle and the S-phase and the probability that a newly produced cell will die within the first 10 days do not differ in these two strains. This indicates that genetic regulation of the size of the proliferating population is independent of the genetic regulation of cell death among those newly produced cells. The dynamic changes in the number of labeled cells as revealed by the SSM protocol also indicate that neither single nor repeated daily injections of BUdR accurately measure 'proliferation.'.

Non-NASA Center↗
NASA NTRS1990

Gravity-induced changes in intracellular potentials in elongating cortical cells of mung bean roots

Gravity-induced changes in intracellular potentials in primary roots of 2-day-old mung bean (Vigna mungo L. cv. black matpe) seedlings were investigated using glass microelectrodes held by 3-dimensional hydraulic micro-drives. The electrodes were inserted into outer cortical cells within the elongation zone. Intracellular potentials, angle of root orientation with respect to gravity, and position within the root of the impaled cortical cell were measured simultaneously. Gravistimulation caused intracellular potential changes in cortical cells of the elongation zone. When the roots were oriented vertically, the intracellular potentials of the outer cortical cells (2 mm behind the root apex) were approximately - 115 mV. When the roots were placed horizontally cortical cells on the upper side hyperpolarized to - 154 mV within 30 s while cortical cells on the lower side depolarized to about - 62 mV. This electrical asymmetry did not occur in cells of the maturation zone. Because attempts to insert the electrode into cells of the root cap were unsuccessful, these cells were not measured. The hyperpolarization of cortical cells on the upper side was greatly reduced upon application of N,N'-dicyclohexylcarbodiimide (DCCD), an inhibitor of respiratory energy coupling. When stimulated roots were returned to the vertical, the degree of hyperpolarization of cortical cells on the previous upper side decreased within 30 s and approached that of cortical cells in non-stimulated roots. This cycle of hyperpolarization/loss of hyperpolarization was repeatable at least ten times by alternately turning the root from the vertical to the horizontal and back again. The very short (<30 s) lag period of these electrical changes indicates that they may result from stimulus-perception and transduction within the elongation zone rather than from transmission of a signal from the root cap.

NASA Discipline Plant Biology↗
NASA NTRS2000

Growth and differentiation of human lens epithelial cells in vitro on matrix

PURPOSE: To characterize the growth and maturation of nonimmortalized human lens epithelial (HLE) cells grown in vitro. METHODS: HLE cells, established from 18-week prenatal lenses, were maintained on bovine corneal endothelial (BCE) extracellular matrix (ECM) in medium supplemented with basic fibroblast growth factor (FGF-2). The identity, growth, and differentiation of the cultures were characterized by karyotyping, cell morphology, and growth kinetics studies, reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence, and Western blot analysis. RESULTS: HLE cells had a male, human diploid (2N = 46) karyotype. The population-doubling time of exponentially growing cells was 24 hours. After 15 days in culture, cell morphology changed, and lentoid formation was evident. Reverse transcription-polymerase chain reaction (RT-PCR) indicated expression of alphaA- and betaB2-crystallin, fibroblast growth factor receptor 1 (FGFR1), and major intrinsic protein (MIP26) in exponential growth. Western analyses of protein extracts show positive expression of three immunologically distinct classes of crystallin proteins (alphaA-, alphaB-, and betaB2-crystallin) with time in culture. By Western blot analysis, expression of p57(KIP2), a known marker of terminally differentiated fiber cells, was detectable in exponential cultures, and levels increased after confluence. MIP26 and gamma-crystallin protein expression was detected in confluent cultures, by using immunofluorescence, but not in exponentially growing cells. CONCLUSIONS: HLE cells can be maintained for up to 4 months on ECM derived from BCE cells in medium containing FGF-2. With time in culture, the cells demonstrate morphologic characteristics of, and express protein markers for, lens fiber cell differentiation. This in vitro model will be useful for investigations of radiation-induced cataractogenesis and other studies of lens toxicity.

Non-NASA Center↗
NASA NTRS1998

The cytoskeleton of Drosophila-derived Schneider line-1 and Kc23 cells undergoes significant changes during long-term culture

Insect cell cultures derived from Drosophila melanogaster are increasingly being used as an alternative system to mammalian cell cultures, as they are amenable to genetic manipulation. Although Drosophila cells are an excellent tool for the study of genes and expression of proteins, culture conditions have to be considered in the interpretation of biochemical results. Our studies indicate that significant differences occur in cytoskeletal structure during the long-term culture of the Drosophila-derived cell lines Schneider Line-1 (S1) and Kc23. Scanning, transmission-electron, and immunofluorescence microscopy studies reveal that microfilaments, microtubules, and centrosomes become increasingly different during the culture of these cells from 24 h to 7-14 days. Significant cytoskeletal changes are observed at the cell surface where actin polymerizes into microfilaments, during the elongation of long microvilli. Additionally, long protrusions develop from the cell surface; these protrusions are microtubule-based and establish contact with neighboring cells. In contrast, the microtubule network in the interior of the cells becomes disrupted after four days of culture, resulting in altered transport of mitochondria. Microtubules and centrosomes are also affected in a small percent of cells during cell division, indicating an instability of centrosomes. Thus, the cytoskeletal network of microfilaments, microtubules, and centrosomes is affected in Drosophila cells during long-term culture. This implies that gene regulation and post-translational modifications are probably different under different culture conditions.

NASA Discipline Developmental Biology↗
NASA NTRS1996

The effect of in vivo and in vitro irradiation (25 Gy) on the subsequent in vitro growth of satellite cells

The effect of in vivo and in vitro irradiation on subsequent satellite cell growth, in vitro, was investigated to ascertain the ability of a 25 Gy dose to inhibit satellite cell proliferation. Satellite cells were isolated from the left (irradiated) and right (non-irradiated) Pectoralis thoracicus of two-week-old tom turkeys 16 h (n=3) and seven weeks (n=2) after the left Pectoralis thoracicus had been irradiated (25 Gy). Satellite cells isolated from the irradiated and non-irradiated muscles exhibited similar (P>0.10) in vitro proliferation indicating that a population of satellite cells survived an in vivo dose of 25 Gy. In additional experiments, satellite cell cultures derived from tom turkey Pectoralis thoracicus were irradiated (25 Gy) in vitro. The number of satellite cells did not (P>0.05) increase in irradiated cultures for 134 h following irradiation, while satellite cells in non-irradiated cultures proliferated (P<0.05) over this time. At later time periods, satellite cell number increased (P<0.05) in irradiated cultures indicating that a population of satellite cells survived irradiation. The results of these in vitro experiments suggest that a 25 Gy dose of irradiation does not abolish satellite cell divisions in the turkey Pectoralis thoracicus.

NASA Discipline Musculoskeletal↗
DOE OSTI2021

MicroRNA-7 overexpression positively regulates the CD8+ SP cell development via targeting PIK3R1

Highlights: • MicroRNA-7 overexpression promotes CD8{sup +} SP cell development. • We identified PIK3R1 as a target of miR-7. • The miR-7/PIK3R1 axis regulates CD8{sup +} SP cell development via modulating the balance of AKT/ERK pathways. microRNA-7 (miR-7), a distinct miRNA family member, has been reported to be involved in the biological functions of immune cells. However, the potential role of miR-7 in the CD8{sup +} T cell development remains to be elucidated. In this study, we estimated the potential effects of miR-7 overexpression in the thymic CD8{sup +} SP cell development using miR-7 overexpression mice. Our results showed that compared with those in control wild type (WT) mice, the volume, weight and total cell numbers of thymus in miR-7 overexpression (OE) mice increased significantly. The absolute cell number of CD8{sup +} SP cells in miR-7 OE mice increased and its ability of activation and proliferation enhanced. Futhermore, we clarified that miR-7 overexpression had an intrinsic promote role in CD8{sup +} SP cell development by adoptive cell transfer assay. Mechanistically, the expression level of PIK3R1, a target of miR-7, decreased significantly in CD8{sup +} SP cells of miR-7 OE mice. Moreover, the expression level of phosphorylated (p)-AKT and p-ERK changed inversely and indicating that miR-7 overexpression impaired the balance of AKE and ERK pathways. In summary, our work reveals an essential role of miR-7 in promoting CD8{sup +} SP cell development through the regulation of PIK3R1 and balance of AKT and ERK pathways.

60 APPLIED LIFE SCIENCES↗