Cardano M.,University of Milan |
Diaferia G.R.,Integrated Systems Engineering |
Cattaneo M.,Institute for Genetic and Biomedical Research |
Dessi S.S.,Integrated Systems Engineering |
And 7 more authors.
Journal of Biological Chemistry | Year: 2011
Murine SEL-1L (mSEL-1L) is a key component of the endoplasmic reticulum-associated degradation pathway. It is essential during development as revealed by the multi-organ dysfunction and in uterus lethality occurring in homozygous mSEL-1L-deficient mice. Here we show that mSEL-1L is highly expressed in pluripotent embryonic stem cells and multipotent neural stem cells (NSCs) but silenced in all mature neural derivatives (i.e. astrocytes, oligodendrocytes, and neurons) by mmumiR-183. NSCs derived from homozygous mSEL-1L-deficient embryos (mSEL-1L-/- NSCs) fail to proliferate in vitro, show a drastic reduction of the Notch effector HES-5, and reveal a significant down-modulation of the early neural progenitor markers PAX-6 and OLIG-2, when compared with the wild type (mSEL-1L+/+ NSCs) counterpart. Furthermore, these cells are almost completely deprived of the neural marker Nestin, display a significant decrease of SOX-2 expression, and rapidly undergo premature astrocytic commitment and apoptosis. The data suggest severe self-renewal defects occurring in these cells probably mediated by misregulation of the Notch signaling. The results reported here denote mSEL-1L as a primitive marker with a possible involvement in the regulation of neural progenitor stemness maintenance and lineage determination. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.
Cardano M.,Vita-Salute San Raffaele University |
Diaferia G.R.,Italian National Cancer Institute |
Falavigna M.,Integrated Systems Engineering |
Spinelli C.C.,Integrated Systems Engineering |
And 3 more authors.
Journal of Histochemistry and Cytochemistry | Year: 2013
Tissue microarray (TMA) and cell microarray (CMA) are two powerful techniques that allow for the immunophenotypical characterization of hundreds of samples simultaneously. In particular, the CMA approach is particularly useful for immunophenotyping new stem cell lines (e.g., cardiac, neural, mesenchymal) using conventional markers, as well as for testing the specificity and the efficacy of newly developed antibodies. We propose the use of a tissue arrayer not only to perform protein expression profiling by immunohistochemistry but also to carry out molecular genetics studies. In fact, starting with several tissues or cell lines, it is possible to obtain the complete signature of each sample, describing the protein, mRNA and microRNA expression, and DNA mutations, or eventually to analyze the epigenetic processes that control protein regulation. Here we show the results obtained using the Galileo CK4500 TMA platform. © The Author(s) 2012.
Baronchelli S.,National Research Council Italy |
La Spada A.,National Research Council Italy |
Conforti P.,University of Milan |
Redaelli S.,University of Milan Bicocca |
And 4 more authors.
Stem Cells and Development | Year: 2015
The potential use of human embryonic stem cells (hESCs) in cell-based therapies points out the critical importance of epigenomic evaluation for cell-based therapies. Specifically, DNA methylation appears to be a crucial player in establishing cell fate commitment and lineage choices. In this study, we report the global changes observed on the CpG islands distributed in promoters, gene bodies, and intergenic regions and the major biochemical pathways and genes involved in methylation changes as H9-hESCs turn into a neuronal culture containing medium-sized spiny striatal neurons (MSNs). Using an ontogeny-recapitulating protocol of striatal neuron differentiation, we analyzed DNA methylation profiles during the conversion from pluripotency to neuropotency up to the acquisition of a mature neuronal phenotype. H9-hESCs changed the methylation pattern both through de novo methylation and hypomethylation of specific gene promoters. Bioinformatic analysis allowed us to identify a panel of striatal-associated genes, which were regulated by DNA methylation and differentially expressed during striatal commitment. Importantly, DNA methylation analysis revealed that H9-hESCs did not acquire methylation-based oncogenic properties after differentiation. Indeed, hypermethylation of cancer-associated genes that characterize transformed cells, such as Polycomb repressive complex-associated genes, was not detected in the neuronal cultures. However, the oncosuppressor gene, BCL2L11, became hypermethylated in H9-hESC-derived mature neurons. Whole-genome DNA methylation profiling could become a technological platform to predict the differentiative potential of hESC-derived cultures and establish further biosafety assessment quality control tools of the cell-based products. © Mary Ann Liebert, Inc. 2015.
Mehta J.P.,Integrated Systems Engineering |
Lavender S.A.,Integrated Systems Engineering |
Lavender S.A.,Ohio State University |
Jagacinski R.J.,Ohio State University |
Sommerich C.M.,Integrated Systems Engineering
Journal of Occupational and Environmental Hygiene | Year: 2015
This study investigated changes in the physiological and behavioral responses to repetitive asymmetric lifting activity after exposure to whole body vibrations. Seventeen healthy volunteers repeatedly lifted a box (15% of lifter's capacity) positioned in front of them at ankle level to a location on their left side at waist level at the rate of 10 lifts/min for a period of 60 minutes. Prior to lifting, participants were seated on a vibrating platform for 60 minutes; in one of the two sessions the platform did not vibrate. Overall, the physiological responses assessed using near-infrared spectroscopy signals for the erector spinae muscles decreased significantly over time during the seating and the lifting tasks (p < 0.001). During repetitive asymmetric lifting, behavioral changes included increases in peak forward bending motion, twisting movement, and three-dimensional movement velocities of the spine the lateral bending movement of the spine and the duration of each lift decreased significantly over the 60 minutes of repetitive lifting. With exposure to whole body vibration, participants twisted farther (p = 0.046) and twisted faster (p = 0.025) these behavioral changes would suggest an increase in back injury risk when repetitive lifting tasks are preceded by whole body vibration exposure. © 2015 Copyright © 2015 JOEH, LLC.
D'Urso D.G.,CNR Institute of Neuroscience |
La Spada A.,CNR Institute of Neuroscience |
Tramonte T.,IRCCS Multimedica |
Rainoldi B.,IRCCS Multimedica |
De Blasio A.,Integrated Systems Engineering
Biopreservation and Biobanking | Year: 2015
In the past decade, the popularity and power of Tissue Microarray (TMA) technology has increased since it provides a method to detect diagnostic and prognostic markers in an array of clinical tissue specimens collected for translational research. TMAs allow for rapid and cost-effective analysis of hundreds of molecular markers at the nucleic acid and protein levels. This technology is particularly useful in the realization of the Human Protein Atlas Project, since it aims to create a reference database of non-redundant human proteins. In this context, it is important to assure the lack of cross-sample contamination due to the repeated use of the same needle in consecutive coring. Here we show that carry-over contamination from one tissue core to another does not occur, reinforcing the accuracy of the TMA technology in the simultaneous testing of multiple bio-samples. © Copyright 2015, Mary Ann Liebert, Inc. 2015.