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https://www.readbyqxmd.com/read/27932699/a-systematic-investigation-of-the-best-buffers-for-use-in-screening-by-maldi-mass-spectrometry
#1
Jessica Chandler, Carl Haslam, Neil Hardy, Melanie Leveridge, Peter Marshall
Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) offers a label-free alternative for the screening of biochemical targets in both 1536- and 6144-assay formats, as well as potentially providing increased sensitivity, reproducibility, and the simultaneous detection of multiple assay components within a specified m/z range. Ion suppression effects are one of the principal limitations reported for MS analysis. Within MALDI-MS screening, it has been identified that certain biochemical components incorporated into the assay (e...
December 8, 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27931511/myod-overexpressed-equine-adipose-derived-stem-cells-enhanced-myogenic-differentiation-potential
#2
Soo-Eun Sung, Meeyul Hwang, Ah-Young Kim, Eun-Mi Lee, Eun-Joo Lee, Su-Kyeong Hwang, Shin-Yoon Kim, Hong-Kyun Kim, Kyu-Shik Jeong
Mesenchymal stem cells could potentially be used in the clinical treatment of muscle disorders and muscle regeneration. Adipose-derived stem cells (ADSCs) can be easily isolated from adipose tissue, as opposed to stem cells of other tissues. We believe that cell therapy using ADSCs could be applied to muscle disorders in horses and other species. We sought to improve the myogenic differentiation potential of equine ADSCs (eqADSCs) using a MyoD lentiviral vector. MyoD lentiviruses were transduced into eqADSCs and selected using puromycin...
November 2016: Cell Transplantation
https://www.readbyqxmd.com/read/27929069/th1-cells-downregulate-connexin-43-gap-junctions-in-astrocytes-via-microglial-activation
#3
Mitsuru Watanabe, Katsuhisa Masaki, Ryo Yamasaki, Jun Kawanokuchi, Hideyuki Takeuchi, Takuya Matsushita, Akio Suzumura, Jun-Ichi Kira
We previously reported early and extensive loss of astrocytic connexin 43 (Cx43) in acute demyelinating lesions of multiple sclerosis (MS) patients. Because it is widely accepted that autoimmune T cells initiate MS lesions, we hypothesized that infiltrating T cells affect Cx43 expression in astrocytes, which contributes to MS lesion formation. Primary mixed glial cell cultures were prepared from newborn mouse brains, and microglia were isolated by anti-CD11b antibody-conjugated magnetic beads. Next, we prepared astrocyte-rich cultures and astrocyte/microglia-mixed cultures...
December 8, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27928705/semi-quantitative-assessments-of-dextran-toxicity-on-corneal-endothelium-conceptual-design-of-a-predictive-algorithm
#4
Filip Filev, Ceprail Oezcan, Jana Feuerstacke, Stephan J Linke, Birgit Wulff, Olaf J C Hellwinkel
Dextran is added to corneal culture medium for at least 8 h prior to transplantation to ensure that the cornea is osmotically dehydrated. It is presumed that dextran has a certain toxic effect on corneal endothelium but the degree and the kinetics of this effect have not been quantified so far. We consider that such data regarding the toxicity of dextran on the corneal endothelium could have an impact on scheduling and logistics of corneal preparation in eye banking. In retrospective statistic analyses, we compared the progress of corneal endothelium (endothelium cell loss per day) of 1334 organ-cultured corneal explants in media with and without dextran...
December 7, 2016: Cell and Tissue Banking
https://www.readbyqxmd.com/read/27927847/preserved-seminiferous-tubule-integrity-with-spermatogonial-survival-and-induction-of-sertoli-and-leydig-cell-maturation-after-long-term-organotypic-culture-of-prepubertal-human-testicular-tissue
#5
F de Michele, J Poels, L Weerens, C Petit, Z Evrard, J Ambroise, D Gruson, C Wyns
STUDY QUESTION: Is an organotypic culture system able to provide the appropriate testicular microenvironment for in-vitro maturation of human immature testicular tissue (ITT)? SUMMARY ANSWER: Our organotypic culture system provided a microenvironment capable of preserving seminiferous tubule (ST) integrity and Leydig cell (LC) functionality and inducing Sertoli cell (SC) maturation. WHAT IS KNOWN ALREADY: Cryopreservation of human ITT is a well-established strategy to preserve fertility in prepubertal boys affected by cancer, with a view for obtaining sperm...
December 6, 2016: Human Reproduction
https://www.readbyqxmd.com/read/27925336/non-exponential-growth-of-mycobacterium-leprae-thai-53-strain-cultured-in-vitro
#6
Kazunobu Amako, Ken-Ichiro Iida, Mitsumasa Saito, Yoshitosi Ogura, Tetsuya Hayashi, Shin-Ichi Yoshida
To determine why cultivation of Mycobacterium leprae in vitro has not been successful, we attempted to culture this bacterium in media supplemented with a variety of biological materials. We observed a slight increase in the number of cells in medium supplemented with human blood plasma and an extract of nude mouse tissue after more than 3 months of cultivation at 30°C. To ascertain whether this increase was real growth, we analyzed the growth by droplet digital PCR. The analysis had the following two results: the slow increase in the copy number of cell-associated DNA and the release of a large amount of DNA into the culture medium from bacterial cells during cultivation...
December 7, 2016: Microbiology and Immunology
https://www.readbyqxmd.com/read/27925198/cancer-cell-plasticity-rapid-reversal-of-chemosensitivity-and-expression-of-stemness-markers-in-lung-and-breast-cancer-tumorspheres
#7
Juan Sebastian Yakisich, Neelam Azad, Vivek Kaushik, Anand Krishnan V Iyer
In cancer cells, the reversible nature of the stemness status in terms of chemoresistance has been poorly characterized. In this study, we have simulated on cycle of environmental conditions to study such reversibility by first generating floating tumorspheres (FTs) from lung and breast cancer cells by culturing them in serum-free media without the addition of any external mitogenic stimulation, and subsequently (after two weeks) re-incubating them back in serum-containing media to simulate routine culture conditions (RCCs)...
December 7, 2016: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/27924610/growing-adipose-derived-stem-cells-under-serum-free-conditions
#8
Diogo Godoy Zanicotti, Dawn E Coates
Growing adipose-derived stem cells (ADSC) in serum-free conditions is important as it represents a way of expanding multipotent cells in a clinical grade medium. Most cultured ADSC are expanded and tested in serum-containing media, which can pose significant health risks if these cells were used in clinical applications. Moreover, cells grown in serum-free conditions behave significantly different than those cultured in serum-containing media. Here, we present a technique to culture adipose-derived stem cells in serum-free conditions...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27924292/data-on-melanin-production-in-b16f1-melanoma-cells-in-the-presence-of-emu-oil
#9
Minoru Ito, Kazuhiro Minami, Yoshimasa Sagane, Toshihiro Watanabe, Koichi Niwa
Here, we present data on the effects of emu oil, obtained from emu (Dromaius novaehollandiae) fat deposits, on melanogenesis in B16F1 murine melanoma cells. The cells were cultured in media containing different concentrations of emu oil, and the melanin content of these cells was measured using a microplate reader. Next, melanin content was measured for cells cultured with α-melanocyte-stimulating hormone. This article reports the different melanin contents as μg melanin/mg cellular protein, by using bar graphs with error bars...
December 2016: Data in Brief
https://www.readbyqxmd.com/read/27924203/the-effects-of-etomidate-and-midazolam-on-adipose-tissue-derived-mesenchymal-stem-cell-proliferation
#10
Ho Park, Kyoungho Ryu, Yun-Hong Kim, Won-Jun Choi, Dongchan Ko
BACKGROUND: Stem cell therapy using adipose tissue-derived mesenchymal stem cells (ADSCs), which are capable of multipotent differentiation, is currently being investigated in the field of tissue regeneration and the treatment of patients in intensive care units. It is known that type-A γ-aminobutyric acid (GABAA) receptor activity has an influence on stem cell proliferation. Thus, we investigated the effects of the clinically available GABAA receptor agonists, etomidate and midazolam, on ADSC proliferation measured by the cell counting kit-8 assay...
December 2016: Korean Journal of Anesthesiology
https://www.readbyqxmd.com/read/27921242/human-mesenchymal-stem-cells-from-adipose-tissue-differentiated-into-neuronal-or-glial-phenotype-express-different-aquaporins
#11
Rosanna Avola, Adriana Carol Eleonora Graziano, Giovanna Pannuzzo, Venera Cardile
Aquaporins (AQPs) are 13 integral membrane proteins that provide selective pores for the rapid movement of water and other uncharged solutes, across cell membranes. Recently, AQPs have been focused for their role in production, circulation, and homeostasis of the cerebrospinal fluid and their importance in several human diseases is becoming clear. This study investigated the time course (0, 14, and 28 days) of AQP1, 4, 7, 8, and 9 during the neural differentiation of human mesenchymal stem cells (MSCs) from adipose tissue (AT)...
December 5, 2016: Molecular Neurobiology
https://www.readbyqxmd.com/read/27919687/cardiomyocyte-released-factors-stimulate-oligodendrocyte-precursor-cells-proliferation
#12
Mariko Kuroda, Rieko Muramatsu, Toshihide Yamashita
The heart produces multiple diffusible factors that are involved in a number of physiological processes, but the action of these factors on the central nervous system is not well understood. In this study, we found that one or more factors released by cardiomyocytes promote oligodendrocyte precursor cell (OPC) proliferation in vitro. Mouse OPCs co-cultured with mouse cardiomyocytes showed higher proliferative ability than OPCs cultured alone. In addition, cardiomyocyte-conditioned media was sufficient to promote OPC proliferation...
December 2, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27919266/arrest-at-the-diplotene-stage-of-meiotic-prophase-i-is-delayed-by-progesterone-but-is-not-required-for-primordial-follicle-formation-in-mice
#13
Sudipta Dutta, Deion M Burks, Melissa E Pepling
BACKGROUND: In mammalian females, reproductive capacity is determined by the size of the primordial follicle pool. During embryogenesis, oogonia divide mitotically but cytokinesis is incomplete so oogonia remain connected in germ cell cysts. Oogonia begin to enter meiosis at 13.5 days postcoitum in the mouse and over several days, oocytes progress through the stages of meiotic prophase I arresting in the diplotene stage. Concurrently, germ cell cysts break apart and individual oocytes become surrounded by granulosa cells forming primordial follicles...
December 5, 2016: Reproductive Biology and Endocrinology: RB&E
https://www.readbyqxmd.com/read/27918831/induction-of-chemokine-secretion-and-monocyte-migration-by-human-choroidal-melanocytes-in-response-to-proinflammatory-cytokines
#14
Tina Jehs, Carsten Faber, Maja S Udsen, Martine J Jager, Simon J Clark, Mogens H Nissen
Purpose: To determine to which extent inflammatory cytokines affect chemokine secretion by primary human choroidal melanocytes (HCMs), their capacity to attract monocytes, and whether HCMs are able to influence the proliferation of activated T cells. Methods: Primary cultures of HCMs were established from eyes of 13 donors. Human choroidal melanocytes were stimulated with IFN-γ and TNF-α or with supernatant from activated T cells (T-cell-conditioned media [TCM])...
December 1, 2016: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/27918556/effects-of-fatty-acid-synthase-inhibitors-on-lymphatic-vessels-an-in-vitro-and-in-vivo-study-in-a-melanoma-model
#15
Débora C Bastos, Jenny Paupert, Catherine Maillard, Fabiana Seguin, Marco A Carvalho, Michelle Agostini, Ricardo D Coletta, Agnès Noël, Edgard Graner
Fatty acid synthase (FASN) is responsible for the endogenous production of fatty acids from acetyl-CoA and malonyl-CoA. Its overexpression is associated with poor prognosis in human cancers including melanomas. Our group has previously shown that the inhibition of FASN with orlistat reduces spontaneous lymphatic metastasis in experimental B16-F10 melanomas, which is a consequence, at least in part, of the reduction of proliferation and induction of apoptosis. Here, we sought to investigate the effects of pharmacological FASN inhibition on lymphatic vessels by using cell culture and mouse models...
December 5, 2016: Laboratory Investigation; a Journal of Technical Methods and Pathology
https://www.readbyqxmd.com/read/27918077/type-2-diabetes-inhibited-human-mesenchymal-stem-cells-angiogenic-response-by-over-activity-of-the-autophagic-pathway
#16
Aysa Rezabakhsh, Omid Cheraghi, Alireza Nourazarian, Mehdi Hassanpour, Masoumeh Kazemi, Shahrooz Ghaderi, Esmaeil Faraji, Reza Rahbarghazi, Çığır Biray Avci, Bakiye Goker Bagca, Alireza Garjani
The current study aimed to address the impact of serum from type 2 diabetes patients on the angiogenic properties of human bone marrow mesenchymal stem cells and its relationship to autophagy signaling. Human primary stem cells were enriched and incubated with serum from diabetic and normal subjects for 7 days. Compared to data from the control group, diabetic serum was found to induce a higher cellular death rate (p < 0.001) and apoptotic changes (p < 0.01). We also showed that diabetic condition significantly abolished angiogenesis tube formation on Matrigel substrate, decreased cell chemotaxis (p < 0...
December 5, 2016: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/27918018/a-planar-model-of-the-vessel-wall-from-cellularized-collagen-scaffolds-focus-on-cell-matrix-interactions-in-mono-bi-and-tri-culture-models
#17
Caroline Loy, Sébastien Meghezi, Lucie Lévesque, Daniele Pezzoli, Heena Kumra, Dieter Reinhardt, Jayachandran N Kizhakkedathu, Diego Mantovani
The acquisition of new thorough knowledge on the interactions existing between vascular cells would represent a step forward in the engineering of vascular tissues. In this light, herein we designed a physiological-like tri-culture in vitro vascular wall model using a planar cellularized collagen gel as the scaffold. The model can be obtained in 24 h and features multi-layered hierarchical organization composed of a fibroblast-containing adventitia-like layer, a media-like layer populated by smooth muscle cells and an intima-like endothelial cell monolayer...
December 5, 2016: Biomaterials Science
https://www.readbyqxmd.com/read/27916562/cryobiological-parameters-of-multipotent-stromal-cells-obtained-from-different-sources
#18
L Lauterboeck, W F Wolkers, B Glasmacher
Stem cells are important for regenerative medicine mainly due to their multilineage differentiation capacity. However, the cells rapidly loose this capability during culturing. Cryopreservation preserves the differentiation potential of the cells, until they are needed. In this study, specific cell properties of multipotent stromal cells (MSCs), from the common marmoset monkey Callithrix jacchus MSCs derived from amnion (Am) and bone marrow (Bm) were studied in order to predict optimal cooling rates for cryopreservation...
December 1, 2016: Cryobiology
https://www.readbyqxmd.com/read/27916353/toll-like-receptor-responses-are-suppressed-in-trauma-icu-patients
#19
Travis L Holloway, Susannah E Nicholson, Meenakshi Rani, Andrew P Cap, Martin G Schwacha
BACKGROUND: Inflammation and activation of the innate immune system are often associated with traumatic injury and may involve alterations in toll-like receptor (TLR)-mediated responses. METHODS: A prospective observational study was designed and conducted. Twenty-one severely injured (ISS = 16-41) trauma intensive care unit (ICU) patients and six healthy volunteers that served as controls were enrolled. Anticoagulated whole blood was collected at 2-12 d after ICU admission and incubated in the presence of media alone (baseline), zymosan (TLR2 agonist) or lipopolysaccharide (LPS; TLR4 agonist) for 3 h...
November 2016: Journal of Surgical Research
https://www.readbyqxmd.com/read/27914320/biochemical-precursor-effects-on-the-fatty-acid-production-in-cell-suspension-cultures-of-theobroma-cacao-l
#20
O Parra, A M Gallego, A Urrea, L F Rojas, C Correa, L Atehortúa
Cocoa butter (CB) is composed of 96% palmitic, stearic, oleic, linoleic and linolenic fatty acids that are responsible for the hardness, texture and fusion properties of chocolate. Through in vitro plant cell culture it is possible to modify CB lipid profiles and to study the fatty acid biosynthesis pathway on a subcellular level, evaluating fundamental aspects to enhance in vitro fatty acid production in a specific and controlled way. In this research, culture media was supplemented with acetate, biotin, pyruvate, bicarbonate and glycerol at three different concentrations and the effects on the biomass production (g/L), cell viability, and fatty acids profile and production was evaluated in in vitro cell suspensions culture...
November 25, 2016: Plant Physiology and Biochemistry: PPB
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