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Research Containing: stem cells

The SCD – Stem Cell Differentiation ESA Project: Preparatory Work for the Spaceflight Mission

by cfynanon 22 August 2016in Biology & Biotechnology No comment

Due to spaceflight, astronauts experience serious, weightlessness-induced bone loss because of an unbalanced process of bone remodeling that involves bone marrow mes- enchymal stem cells (BMSCs), as well as osteoblasts, osteo- cytes, and osteoclasts. The effects of microgravity on osteo- cells have been extensively studied, but it is only recently that consideration has been given to the role of BMSCs. Pre- vious researches indicated that human BMSCs cultured in simulated microgravity (sim-μg) alter their proliferation and differentiation. The spaceflight opportunities for biomedical experiments are rare and suffer from a number of opera- tive constraints that could bias the validity of the experiment itself, but remain a unique opportunity to confirm and explain the effects due to microgravity, that are only par- tially activated/detectable in simulated conditions. For this reason, we carefully prepared the SCD – STEM CELLS DIFFERENTIATION experiment, selected by the European Space Agency (ESA) and now on the International Space Station (ISS). Here we present the preparatory studies per- formed on ground to adapt the project to the spaceflight constraints in terms of culture conditions, fixation and stor- age of human BMSCs in space aiming at satisfying the biological requirements mandatory to retrieve suitable sam- ples for post-flight analyses. We expect to understand better the molecular mechanisms governing human BMSC growth and differentiation hoping to outline new countermeasures against astronaut bone loss.

Related URLs:
http://link.springer.com/article/10.1007/s12217-015-9466-5

Genetic and Apoptotic Changes in Lungs of Mice Flown on the STS-135 Mission in Space.

by cfynanon 22 August 2016in Biology & Biotechnology No comment

AIM: The goal of the study was to evaluate changes in lung status due to spaceflight stressors that include radiation above levels found on Earth.;MATERIALS AND METHODS: Within hours after return from a 13-day mission in space onboard the Space Shuttle Atlantis, C57BL/6 mice (FLT group) were euthanized; mice housed on the ground in similar animal enclosure modules served as controls (AEM group). Lung tissue was collected to evaluate the expression of genes related to extracellular matrix (ECM)/adhesion and stem cell signaling. Pathway analysis was also performed. In addition, immunohistochemistry for stem cell antigen-1 (SCA-1), the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay for apoptosis, and staining for histological characteristics were performed.;RESULTS: There were 18/168 genes significantly modulated in lungs from the FLT group (p<0.05 vs. AEM); 17 of these were up-regulated and one was down-regulated. The greatest effect, namely a 5.14-fold increase, was observed on Spock1 (also known as Spark/osteonectin), encoding a multi-functional protein that has anti-adhesive effects, inhibits cell proliferation and regulates activity of certain growth factors. Additional genes with increased expression were cadherin 3 (Cdh3), collagen, type V, alpha 1 (Col5a1), integrin alpha 5 (Itga5), laminin, gamma 1 (Lamc1), matrix metallopeptidase 14 (Mmp14), neural cell adhesion molecule 1 (Ncam1), transforming growth factor, beta induced (Tgfbi), thrombospondin 1 (Thbs1), Thbs2, versican (Vcan), fibroblast growth factor receptor 1 (Fgfr1), frizzled homolog 6 (Fzd6), nicastrin (Ncstn), nuclear factor of activated T-cells, cytoplasmic, calcineurin-dependent 4 (Nfatc4), notch gene homolog 4 (Notch4) and vang-like 2 (Vangl2). The down-regulated gene was Mmp13. Staining for SCA-1 protein showed strong signal intensity in bronchiolar epithelial cells of FLT mice (p<0.05 vs. AEM). TUNEL positivity was also significantly higher in the FLT mice (p<0.05 vs. AEM), but no consistent histological differences were noted. CONCLUSION: The results demonstrate that spaceflight-related stress had a significant impact on lung integrity, indicative of tissue injury and remodeling. Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/26130787

Microgravity Reduces the Differentiation and Regenerative Potential of Embryonic Stem Cells

by cfynanon 22 August 2016in Biology & Biotechnology No comment

Mechanical unloading in microgravity is thought to induce tissue degeneration by various mechanisms, including inhibition of regenerative stem cell differentiation. To address this hypothesis, we investigated the effects of microgravity on early lineage commitment of mouse embryonic stem cells (mESCs) using the embryoid body (EB) model of tissue differentiation. We found that exposure to microgravity for 15 days inhibits mESC differentiation and expression of terminal germ layer lineage markers in EBs. Additionally, microgravity-unloaded EBs retained stem cell self-renewal markers, suggesting that mechanical loading at Earth’s gravity is required for normal differentiation of mESCs. Finally, cells recovered from microgravity-unloaded EBs and then cultured at Earth’s gravity showed greater stemness, differentiating more readily into contractile cardiomyocyte colonies. These results indicate that mechanical unloading of stem cells in microgravity inhibits their differentiation and preserves stemness, possibly providing a cellular mechanistic basis for the inhibition of tissue regeneration in space and in disuse conditions on earth.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/26414276

Decrease in the number of progenitors of fibroblasts (CFUf) in bone marrow of rats after a 14-day flight onboard the Cosmos-2044 biosatellite

by cfynanon 9 June 2015in Biology & Biotechnology No comment

A decrease in the number of progenitors of fibroblasts (CFUf) was found in bone marrow of rats that underwent a 14-day flight in the state of weightlessness onboard the Cosmos-2044 biosatellite, immediately after flight, when compared with rats maintained in control conditions of terrestrial gravitation. These changes may be explained by the action of specific factors of microgravitation.

Related URLs:
http://ovidsp.ovid.com/ovidweb.cgi?T=JS&CSC=Y&NEWS=N&PAGE=fulltext&D=med3&AN=2257939
http://sfxhosted.exlibrisgroup.com/mayo?sid=OVID:medline&id=pmid:2257939&id=doi:&issn=0015-5500&isbn=&volume=36&issue=3&spage=194&pages=194-7&date=1990&title=Folia+Biologica&atitle=Decrease+in+the+number+of+progenitors+of+fibroblasts+%28CFUf%29+in+bone+marrow+of+rats+after+a+14-day+flight+onboard+the+Cosmos-2044+biosatellite.&aulast=Vacek&pid=%3Cauthor%3EVacek+A%3C%2Fauthor%3E&%3CAN%3E2257939%3C%2FAN%3E

The application of low shear modeled microgravity to 3-D cell biology and tissue engineering

by cfynanon 9 June 2015in Biology & Biotechnology No comment

The practice of cell culture has been virtually unchanged for 100 years. Until recently, life scientists have had to content themselves with two-dimensional cell culture technology. Clearly, living creatures are not constructed in two dimensions and thus it has become widely recognized that in vitro culture systems must become three dimensional to correctly model in vivo biology. Attempts to modify conventional 2-D culture technology to accommodate 3-D cell growth such as embedding cells in extracellular matrix have demonstrated the superiority of concept. Nevertheless, there are serious drawbacks to this approach including limited mass transport and lack of scalability. Recently, a new cell culture technology developed at NASA to study the effects of microgravity on cells has emerged to solve many of the problems of 3-D cell culture. The technology, the Rotating Wall Vessel (RWV) is a single axis clinostat consisting of a fluid-filled, cylindrical, horizontally rotating culture vessel. Cells placed in this environment are suspended by the resolution of the gravitational, centrifugal and Coriolis forces with extremely low mechanical shear. These conditions, which have been called “low shear modeled microgravity”, enable cells to assemble into tissue-like aggregates with high mass transport of nutrients, oxygen and wastes. Examples of the use of the RWV for basic cell biology research and tissue engineering applications are discussed.

Related URLs:
http://ovidsp.ovid.com/ovidweb.cgi?T=JS&CSC=Y&NEWS=N&PAGE=fulltext&D=emed8&AN=2008312724
http://sfxhosted.exlibrisgroup.com/mayo?sid=OVID:embase&id=pmid:&id=doi:10.1016%2FS1387-2656%252808%252900011-2&issn=1387-2656&isbn=9780444532268&volume=14&issue=&spage=275&pages=275-296&date=2008&title=Biotechnology+Annual+Review&atitle=The+application+of+low+shear+modeled+microgravity+to+3-D+cell+biology+and+tissue+engineering&aulast=Navran&pid=%3Cauthor%3ENavran+S.%3C%2Fauthor%3E&%3CAN%3E2008312724%3C%2FAN%3E

Effects of microgravity and increased gravity on bone marrow of rats

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Astronauts have a reduction in their red cell mass when exposed to microgravity. This is probably mainly due to a physiological response to decreased energy requirements. Further studies of erythropoiesis were carried out in microgravity on rats flown on Soviet Biosatellite 2044 and in hypergravity by centrifugation at 2G. Studies included: bone marrow cell differential counts, clonal studies of RBC colony formation, and plasma erythropoietin determinations. In the bone marrow of Cosmos flight animals there was a slight increase in granulocytic cells and in centrifuged animals, a slight decrease in the percentage of erythroid cells which led to an increased M:E ratio. The bone marrow cells of flight and centrifuged rats responded to erythropoietin. Cosmos flight animals' cells formed fewer CFU-E than the controls but this was reversed in the centrifuge studies. There were no essential differences in the erythropoietin levels of test groups as compared to control groups.

Related URLs:
http://ovidsp.ovid.com/ovidweb.cgi?T=JS&CSC=Y&NEWS=N&PAGE=fulltext&D=med3&AN=7980333
http://sfxhosted.exlibrisgroup.com/mayo?sid=OVID:medline&id=pmid:7980333&id=doi:&issn=0095-6562&isbn=&volume=65&issue=8&spage=730&pages=730-5&date=1994&title=Aviation+Space+%26+Environmental+Medicine&atitle=Effects+of+microgravity+and+increased+gravity+on+bone+marrow+of+rats.&aulast=Lange&pid=%3Cauthor%3ELange+RD%3C%2Fauthor%3E&%3CAN%3E7980333%3C%2FAN%3E

Effect of simulated weightlessness on osteoprogenitor cell number and proliferation in young and adult rats

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Experiments with rats flown in space or hind limb unloaded (HU) indicate that bone loss in both conditions is associated with a decrease in bone volume and osteoblast surface in cancellous and cortical bone. We hypothesize that the decrease in osteoblastic bone formation and osteoblast surface is related to a decrease in the number of osteoprogenitors and/or decreased proliferation of their progeny. We tested this hypothesis by evaluating the effect of 14 days of HU on the number of osteoprogenitors (osteoblast colony forming units; CFU-O), fibroblastic colony forming units (CFU-F), and alkaline phosphatase-positive CFU (CFU-AP) in cell populations derived from the proximal femur (unloaded) and the proximal humerus (normally loaded) in 6-week-old and 6-month-old rats. To confirm the effect of unloading on bone volume and structure, static histomorphometric parameters were measured in the proximal tibial metaphysis. Effects of HU on proliferation of osteoprogenitors were evaluated by measuring the size of CFU-O. HU did not affect the total number of progenitors (CFU-F) in young or adult rats in any of the cell populations. In femoral populations of young rats, HU decreased CFU-O by 71.0% and mean colony size was reduced by 20%. HU decreased CFU-AP by 31.3%. As expected, no changes in CFU-O or CFU-AP were seen in cell populations from the humerus. In femoral cell populations of adult rats, HU decreased CFU-O and CFU-AP by 16.6% and 36.6%, respectively. Again, no effects were seen in cell populations from the humerus. In 6-week-old rats, there was a greater decrease in bone volume, osteoblast number, and osteoblast surface in the proximal tibial metaphysis than that observed in adult rats. Both trabecular thickness and trabecular number were decreased in young rats but remained unaffected in adults. Neither osteoclast number nor surface was affected by unloading. Our results show that the HU-induced decrease in the number of osteoprogenitors observed in vitro parallels the effects of HU on bone volume and osteoblast number in young and old rats in vivo, suggesting that the two may be interdependent. HU also reduced CFU-O colony size in femoral populations indicating a diminished proliferative capacity of osteoblastic colonies.

Related URLs:
http://ovidsp.ovid.com/ovidweb.cgi?T=JS&CSC=Y&NEWS=N&PAGE=fulltext&D=med4&AN=15664015
http://sfxhosted.exlibrisgroup.com/mayo?sid=OVID:medline&id=pmid:15664015&id=doi:&issn=8756-3282&isbn=&volume=36&issue=1&spage=173&pages=173-83&date=2005&title=Bone&atitle=Effect+of+simulated+weightlessness+on+osteoprogenitor+cell+number+and+proliferation+in+young+and+adult+rats.&aulast=Basso&pid=%3Cauthor%3EBasso+N%3C%2Fauthor%3E&%3CAN%3E15664015%3C%2FAN%3E

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