Manuela Monti
@unipv.it
Scopus Publications
Scopus Publications
Silvia Pisani, Enrica Chiesa, Ida Genta, Rossella Dorati, Marilena Gregorini, Maria Antonietta Grignano, Marina Ramus, Gabriele Ceccarelli, Stefania Croce, Chiara Valsecchi,et al.
MDPI AG
This work focuses on formulating liposomes to be used in isolated kidney dynamic machine perfusion in hypothermic conditions as drug delivery systems to improve preservation of transplantable organs. The need mainly arises from use of kidneys from marginal donors for transplantation that are more exposed to ischemic/reperfusion injury compared to those from standard donors. Two liposome preparation techniques, thin film hydration and microfluidic techniques, are explored for formulating liposomes loaded with two model proteins, myoglobin and bovine serum albumin. The protein-loaded liposomes are characterized for their size by DLS and morphology by TEM. Protein releases from the liposomes are tested in PERF-GEN perfusion fluid, 4 °C, and compared to the in vitro protein release in PBS, 37 °C. Fluorescent liposome uptake is analyzed by fluorescent microscope in vitro on epithelial tubular renal cell cultures and ex vivo on isolated pig kidney in hypothermic perfusion conditions. The results show that microfluidics are a superior technique for obtaining reproducible spherical liposomes with suitable size below 200 nm. Protein encapsulation efficiency is affected by its molecular weight and isoelectric point. Lowering incubation temperature slows down the proteins release; the perfusion fluid significantly affects the release of proteins sensitive to ionic media (such as BSA). Liposomes are taken up by epithelial tubular renal cells in two hours’ incubation time.
Pierangela Grignani, Alessandro Manfredi, Maria Cristina Monti, Matteo Moretti, Luca Morini, Silvia Damiana Visonà, Paolo Fattorini, and Carlo Previderè
Wiley
Abstract The collection of liquid biological matrices onto paper cards (dried matrix spots [DMS]) is becoming an alternative sampling strategy. The stability over time of molecules of interest for therapeutic, sport drug monitoring, and forensic toxicology on DMS has been recently investigated representing a reliable alternative to conventional analytical techniques. When a tampering of a urine sample in drug monitoring or doping control cases is suspected, it could be relevant to know whether genetic profiles useful for individual identification could be generated from urine samples spotted onto paper (dried urine spot [DUS]). To understand the influence of sex, storage conditions, and time on the quality and quantity of the DNA, five female and ten male urine samples were dispensed onto Whatman 903 paper and sampled after different storage conditions over time, from 1 to 12 weeks. Direct PCR was performed starting from 2‐mm punches collected from each spot amplifying a panel of markers useful for individual identification. The female DUS stored in different conditions produced genetic profiles fully matching the reference samples. The same result was obtained for the male DUS but using urine 30X concentrated by centrifugation instead of the original samples. Our data show that this approach is valid for genetic individual identification of urine samples spotted onto paper cards up to 12 weeks after deposition and could be easily incorporated in anti‐doping or drug screening protocols to help on the suspicion of evidence tampering or to solve questions on the reliability of samples collection.
Alessandra Rodriguez y Baena, Andrea Casasco, and Manuela Monti
Springer Science and Business Media LLC
AbstractOne of the most exciting advances in life science research is the development of 3D cell culture systems to obtain complex structures called organoids and spheroids. These 3D cultures closely mimic in vivo conditions, where cells can grow and interact with their surroundings. This allows us to better study the spatio-temporal dynamics of organogenesis and organ function. Furthermore, physiologically relevant organoids cultures can be used for basic research, medical research, and drug discovery. Although most of the research thus far focuses on the development of heart, liver, kidney, and brain organoids, to name a few, most recently, these structures were obtained using dental stem cells to study in vitro tooth regeneration. This review aims to present the most up-to-date research showing how dental stem cells can be grown on specific biomaterials to induce their differentiation in 3D. The possibility of combining engineering and biology principles to replicate and/or increase tissue function has been an emerging and exciting field in medicine. The use of this methodology in dentistry has already yielded many interesting results paving the way for the improvement of dental care and successful therapies. Graphical abstract
Manuela Monti, CarloAlberto Redi, and Ernesto Capanna
PAGEPress Publications
In this paper, we report genome size (GS) values for nine cockroaches (order Blattodea, families Blattidae, Blaberidae and Ectobiidae, ex Blattelidae,), three of which are original additions to the ten already present in the GS database: the death’s head roach (Blaberus craniifer), the Surinam cockroach (Pycnoscelus surinamensis) and the Madeira cockroach (Leucophaea maderae). Regarding the American cockroach (Periplaneta americana), the GS database contains two contrasting values (2.72 vs 3.41 pg); likely, the 2.72 pg value is the correct one as it is strikingly similar to our sperm DNA content evaluation (2.80 ± 0.11 pg). Also, we suggest halving the published GS of the Argentine cockroach Blaptica dubia and the spotted cockroach (the gray cockroach) Nauphoeta cinerea discussing (i) the occurrence of a correlation between increasing 2N chromosome number and GS within the order Blattodea; and (ii) the possible occurrence of a polyploidization phenomenon doubling a basic GS of 0.58 pg of some termite families (superfamily Blattoidea, epifamily Termitoidae).
Manuela Monti
PAGEPress Publications
This book focuses on a new category of stem cells derived from perinatal tissue, including amniotic fluid, fetal membranes, umbilical cord, and placental tissue....
M. Monti
This book focuses on a new category of stem cells derived from perinatal tissue, including amniotic fluid, fetal membranes, umbilical cord, and placental tissue. Thirteen chapters overview the biology, production, and the use of perinatal stem cells in translational medicine. They are a particular type of stem cells similar to embryonic and adult stem cells and endowed with multipotent phenotypes. As well known, the significantly high differentiative potential of embryonic stem cells, tied to the ethical concerns they raise with the risk of teratoma formation, limits their use in pre-clinical and clinical studies. On the other hand, both somatic stem cells and induced pluripotent stem cells have limitations and disadvantages. Induced pluripotent stem cells may be at risk for epigenetic abnormalities and integration of viral genomes. Somatic stem cells (i.e., the most used cells in regenerative medicine) can be difficult to isolate (and this sometimes becomes a limiting factor) and may have a limited proliferative capacity. Fortunately, new shreds of evidence show that related perinatal tissues (umbilical cord, placenta, and both membrane and amniotic fluid) are new sources of stem cells, known as perinatal stem cells. The latter are classified according to their anatomical origin and divide into three groups: stem cells of the amniotic fluid, placenta, and umbilical cord; moreover, the stem cells of the placenta can be divided into three further main subgroups based on distinct anatomical districts from which they can be isolated (amnion, villi, and blood) while the umbilical cord stem cells are known as cord blood and Wharton’s jelly’s stem cells. Two main biological features make these cells attractive in regenerative medicine. 1. Their differentiation potential lies between embryonic and somatic stem cells according to the stages of development they derive: the umbilical cord blood, amniotic fluid, placenta, postgestational maternal peripheral blood. 2. Their remarkable immunomodulatory properties modulate various cells of the innate and adaptive immune systems, which can expand the applications of stem cells derived from perinatal tissues, thus opening up new therapeutic strategies. These biological features make perinatal tissue-derived stem cells the promise El Dorado for future cellular therapies since they are free of any ethical or biological constraints usually bundled to the other categories (i.e., embryonic and induced pluripotent stem cells). Studying the genetic stability and stemness of perinatal stem cells will undoubtedly advance our understanding of the molecular basis of stem cell pluripotency and plasticity. As I scrolled through the chapters, I discovered the fascinating world of new reagents provided by the perinatal stem cells: the secretomes they produce. It is well known that another look into the future of stem cell therapies comes from the use of stem cell derivatives such as stem cell extracts and exosomes. These soluble factors (proteins) exert paracrine activity in various biological processes such as wound healing, inflammatory response, angiogenesis, and cell proliferation, to name a few. Secretomes produced in conditioned media can be manufactured and then used as protein drugs to treat various diseases in the near future. The exosomes (nanovesicles containing proteins, RNA and DNA) present in the secretomes produced by mesenchymal stem cells (from perinatal tissues) have already been active in regenerative processes and modulating angiogenic effects. Several colored figures, tables, and a very rich bibliography enrich each chapter that composes this book. The reader draws essential information from this reading: mesenchymal stem cells derived from perinatal tissues are already effective in treating the nervous system, cardiovascular, inflammatory, liver diseases, diabetes mellitus, and kidney regeneration. The book offers detailed insights into the sources of perinatal stem cells and how to derive, multiply and store them. It is intended for researchers in stem cell biology, tissue engineering, regenerative medicine, and students in developmental/stem cell biology. Usually discarded as medical waste, such a new and versatile source of stem cells will allow new technologies to meet unmet therapeutic needs. One of the essential needs will likely involve developing effective therapies for the treatment of severe COVID-19: mesenchymal stem cells can inhibit the onset of a cytokine storm and exercise pulmonary fibrosis. Therefore, perinatal tissue-derived mesenchymal stem cells are expected to play an important role, along with massive vaccination campaigns, in restoring human health in the coming years.
Manuela Monti
PAGEPress Publications
Stem cell (SC) biology is a pervasive transdisciplinary research field encompassing any level of life organization (from molecular to morphological), combining different types of techniques (spanning from cellular to molecular).....
Manuela Monti
PAGEPress Publications
The ten chapters of this volume exploit the potentials of stem cells biology in translational medicine in a very clear way also thanks to nice color illustrations together with tricks and suggestions useful to get reproducible results...
Manuela Monti
PAGEPress Publications
Clathrin is one of the interesting “moonlighting proteins” which perform multiple functions relevant to biochemical or biophysical aspects. It can be considered the master regulator of vesicular trafficking being the main player of the Clathrin-Mediated Endocytosis (CME).....
Laura Pandolfi, Vanessa Frangipane, Claudia Bocca, Alessandro Marengo, Erika Tarro Genta, Sara Bozzini, Monica Morosini, Maura D’Amato, Simone Vitulo, Manuela Monti,et al.
MDPI AG
Collagen Tissue Disease–associated Interstitial Lung Fibrosis (CTD-ILDs) and Bronchiolitis Obliterans Syndrome (BOS) represent severe lung fibrogenic disorders, characterized by fibro-proliferation with uncontrolled extracellular matrix deposition. Hyaluronic acid (HA) plays a key role in fibrosis with its specific receptor, CD44, overexpressed by CTD-ILD and BOS cells. The aim is to use HA-liposomes to develop an inhalatory treatment for these diseases. Liposomes with HA of two molecular weights were prepared and characterized. Targeting efficiency was assessed toward CTD-ILD and BOS cells by flow cytometry and confocal microscopy and immune modulation by RT-PCR and ELISA techniques. HA-liposomes were internalized by CTD-ILD and BOS cells expressing CD44, and this effect increased with higher HA MW. In THP-1 cells, HA-liposomes decreased pro-inflammatory cytokines IL-1β, IL-12, and anti-fibrotic VEGF transcripts but increased TGF-β mRNA. However, upon analyzing TGF-β release from healthy donors-derived monocytes, we found liposomes did not alter the release of active pro-fibrotic cytokine. All liposomes induced mild activation of neutrophils regardless of the presence of HA. HA liposomes could be also applied for lung fibrotic diseases, being endowed with low pro-inflammatory activity, and results confirmed that higher MW HA are associated to an increased targeting efficiency for CD44 expressing LFs-derived from BOS and CTD-ILD patients.
Manuela Monti
PAGEPress Publications
As the title suggests, this book presents several techniques to study cells migration in vivo, in vitro, ex vivo and with different model systems to dissect many of the biochemical and biophysical properties (at both molecular and cellular levels) involved in the dynamics of migration and cell-to-cell communication...
Emanuela Cova, Laura Pandolfi, Miriam Colombo, Vanessa Frangipane, Simona Inghilleri, Monica Morosini, Simona Mrakic-Sposta, Sarah Moretti, Manuela Monti, Ymera Pignochino,et al.
Informa UK Limited
Purpose Malignant pleural mesothelioma (MPM) is an aggressive tumor characterized by poor prognosis. Its incidence is steadily increasing due to widespread asbestos exposure. There is still no effective therapy for MPM. Pemetrexed (Pe) is one of the few chemotherapeutic agents approved for advanced-stage disease, although the objective response to the drug is limited. The use of gold nanoparticles (GNPs) as a drug delivery system promises several advantages, including specific targeting of malignant cells, with increased intracellular drug accumulation and reduced systemic toxicity, and, in the case of MPM, direct treatment administration into the pleural space. This study aims at exploring CD146 as a potential MPM cell-specific target for engineered Pe-loaded GNPs and to assess their effectiveness in inhibiting MPM cell line growth. Methods MPM cell lines and primary cultures obtained by pleural effusions from MPM patients were assayed for CD146 expression by flow cytometry. Internalization by MPM cell lines of fluorescent dye-marked GNPs decorated with a monoclonal anti CD146 coated GNPs (GNP-HC) was proven by confocal microscopy. The effects of anti CD146 coated GNPs loaded with Pe (GNP-HCPe) on MPM cell lines were evaluated by cell cycle (flow cytometry), viability (MTT test), clonogenic capacity (soft agar assay), ROS production (electric paramagnetic resonance), motility (wound healing assay), and apoptosis (flow cytometry). Results GNP-HC were selectively uptaken by MPM cells within 1 hour. MPM cell lines were blocked in the S cell cycle phase in the presence of GNP-HCPe. Both cell viability and motility were significantly affected by nanoparticle treatment compared to Pe. Apoptotic rate and ROS production were significantly higher in the presence of nanoparticles. Clonogenic capacity was completely inhibited following nanoparticle internalization. Conclusion GNP-HCPe treatment displays in vitro antineoplastic action and is more effective than Pe alone in inhibiting MPM cell line malignant phenotype. The innovative use of specifically targeted GNPs opens the perspective of local intrapleural administration to avoid normal cell toxicity and enhance chemotherapy efficacy.
Manuela Monti
PAGEPress Publications
The Springer Protocols series “Methods in Molecular Biology” has published its 1818th volume which is entirely devoted to the development of the female gamete: the oocyte. 
Marianna Longo, Michele Boiani, CarloAlberto Redi, and Manuela Monti
PAGEPress Publications
Cytoplasmic lattices are important regulators of oocyte maturation. They store components of the protein synthesis machinery including ribosomes and, among others, they are involved in the regulation of microtubule dynamics in both mouse and human. Cytoplasmic lattices undergo dramatic reorganizations at crucial stages of oocyte maturation, where they are abundantly present in the cytoplasm of developmentally competent oocytes named SN (Surrounded Nucleolus) while they are rare in the cytoplasm of 2-cell stage-arresting NSN (Not Surrounded Nucleolus) oocytes, suggestive of a requirement of cytoplasmic lattices for development past the 2-cell stage. Here, to elucidate this requirement, 2-cell mouse embryos derived from SN and NSN oocytes were analyzed by transmission electron microscopy. Contrary to what had been proposed hitherto, cytoplasmic lattices are present in 2-cell embryos derived not only from SN, but also from NSN oocytes, irrespective of the embryo production system (intra cytoplasmic sperm injection, parthenogenesis). Hence our conclusion that cytoplasmic lattices do not count among the factor(s) responsible for the embryo arrest at this crucial stage of development.
Niccolò Bianchi, Marianna Longo, CarloAlberto Redi, and Manuela Monti
Wiley
A gradual restriction in stem cells potency is observed during the different stages of mammalian embryonic development, starting with the zygote, the totipotent stem cell par excellence, up to the blastocyst stage. Stem cells of the inner cell mass are endowed with pluripotent characteristic, being able to form cells of the three germ layers (ectoderm, endoderm, and mesoderm). Pluripotent stem cells retain proliferative capacity throughout the life of an individual, and divide asymmetrically, wherein one cell is the copy of the original stem cell while the other is programmed to begin the differentiation process. In mammals, pluripotent stem cells can also be found in the fetus and in some adult tissues, including the umbilical cord blood
Manuela Monti
PAGEPress Publications
This book presents different aspects of oocytes maturation in several animal models as the interactions oocyte-environment, the specific functions of oocytes molecules and organelles regulated by epigenetic and translational mechanisms and the evolution of maternal factors...
Manuela Monti, Barbara Imberti, Niccolò Bianchi, Anna Pezzotta, Marina Morigi, Claudia Del Fante, Carlo Alberto Redi, and Cesare Perotti
Mary Ann Liebert Inc
Very small embryonic-like cells (VSELs) are a population of very rare pluripotent stem cells isolated in adult murine bone marrow and many other tissues and organs, including umbilical cord blood (UCB). VSEL existence is still not universally accepted by the scientific community, so for this purpose, we sought to investigate whether presumptive VSELs (pVSELs) could be isolated from human UCB with an improved protocol based on the isolation of enriched progenitor cells by depletion of nonprogenitor cells with magnetic separation. Progenitor cells, likely including VSELs, cultured with retinoic acid were able to form dense colonies and cystic embryoid bodies and to differentiate toward the ecto-meso-endoderm lineages as shown by the positivity to specific markers. VSEL differentiative potential toward mesodermal lineage was further demonstrated in vitro upon exposure to an established inductive protocol, which induced the acquisition of renal progenitor cell phenotype. VSEL-derived renal progenitors showed regenerative potential in a cisplatin model of acute kidney injury by restoring renal function and tubular structure through induction of proliferation of endogenous renal cells. The data presented here foster the great debate that surrounds VSELs and, more in general, the existence of cells endowed with pluripotent features in adult tissues. In fact, the possibility to find and isolate subpopulations of cells that fully fit all the criteria utilized to define pluripotency remains, nowadays, almost unproven. Thus, efforts to better characterize the phenotype of these intriguing cells are crucial to understand their possible applications for regenerative and precision medicine purposes.
Manuela Monti, Antonio Graziano, Silvana Rizzo, Cesare Perotti, Claudia Del Fante, Riccardo d'Aquino, Carlo Alberto Redi, and Ruggero Rodriguez y Baena
Wiley
Human population is facing a revolutionary change in the demographic structure with an increasing number of elderly people requiring an unmet need to ensure a smooth aging process and dental care is certainly an important aspect that has to be considered. To date, dentistry has been conservative and the need of transferring the scientific models of regenerative dentistry into clinical practice is becoming a necessity. The aim of this study was to characterize the differentiation commitment (in vitro) and the clinical grafting ability (in vivo) of a population of progenitor stem cells obtained after mechanical digestion of dental pulp with an innovative system recently developed. This approach was successfully used in previous studies to obtain a clinical‐grade ready to use dental pulp fragments that could be grafted in autologous tissues to obtain bone. We are thus showing that micro grafts resulting from mechanical digestion contain stem cells with a mesenchymal phenotype, able to differentiate toward different cell types and to generate new bone in patients. We are providing data for the establishment of standardized and routinely oral surgery approaches, having outlined the cellular properties of human stem cells obtained from the dental pulp. This method can represent a valid tool for both regenerative medicine and tissue engineering purposes not only applicable to the cranio‐maxillofacial region but, likely, to different bone pathologies for a fastening and healing recovering of patients. J. Cell. Physiol. 232: 548–555, 2017. © 2016 Wiley Periodicals, Inc.
Manuela Monti, Alberto Calligaro, Barry Behr, Renee Rejo Pera, Carlo Alberto Redi, and Mark Wossidlo
PAGEPress Publications
In vivo maturation (IVM) of human oocytes is a technique used to increase the number of usable oocytes for in vitro fertilization (IVF) and represents a necessity for women with different ovarian pathologies. During IVM the oocytes progress from the germinal vesicle stage (GV) through the metaphase II and during this journey both nuclear and cytoplasmic rearrangements must be obtained to increase the probability to get viable and healthy zygotes/embryos after IVF. As the successful clinical outcomes of this technique are a reality, we wanted to investigate the causes behind oocytes maturation arrest. For obvious ethical reasons, we were able to analyze only few human immature oocytes discarded and donated to research by transmission electron microscopy showing that, as in the mouse, they have different chromatin and cytoplasmic organizations both essential for further embryo development.
Kei-ichiro Ishiguro, Manuela Monti, Tomohiko Akiyama, Hiromi Kimura, Nana Chikazawa-Nohtomi, Miki Sakota, Saeko Sato, Carlo Alberto Redi, Shigeru B. H. Ko, and Minoru S. H. Ko
Springer Science and Business Media LLC
Manuela Monti
PAGEPress Publications
By putting together the most advanced evidences supporting the 'gamete and embryo-fetal origins of adult diseases' the two editors, Prof. He-Feng Huang and Prof. Jian-Zhong Sheng (Hangzhou, People’s Republic of China) did a great workk.....
Manuela Monti and Carlo Alberto Redi
MyJove Corporation
This protocol describes a simple and quick method to isolate and characterize mouse antral GV (Germinal Vesicle) oocytes as able (SN, Surrounded Nucleolus) or unable (NSN, Not Surrounded Nucleolus) to develop to the blastocyst stage after in vitro maturation (IVM) and in vitro fertilization (IVF). It makes use of Hoeschst33342 (or any other DNA intercalating dye) able to bind to the heterochromatin of the nucleolus showing a ring in the SN oocytes or not, like in the NSN oocytes. This represents the easiest and quickest way to sort both antral oocytes that can be eventually used for IVM or IVF procedures. Briefly, the protocol consists of the following steps: hormone injection to stimulate follicular growth; isolation of the oocytes at the GV stage from the antral compartment by puncturing the ovary with a sterile needle; preparation of thin glass pipettes for mouth pipetting of the oocytes; sorting of the oocytes with Hoechst33342 prepared at a supravital concentration; IVM, IVF or any other molecular/cellular analysis. Unfortunately there are still few evidences to sort SN and NSN oocytes using less invasive techniques. If and once they will be identified, they could be potentially applied to human assisted reproductive technologies, although with several aspects that should be modified. To date, this technique has potential implications to dramatically increase IVM and IVF successful procedures in both endangered and species with economic interest.
Letizia Trovato, Manuela Monti, Claudia del Fante, Marila Cervio, Milla Lampinen, Lucia Ambrosio, Carlo Alberto Redi, Cesare Perotti, Esko Kankuri, Gennaro Ambrosio,et al.
Wiley
Autologous graft is considered the gold standard of graft materials; however, this approach is still limited due to both small amount of tissue that can be collected and to reduced cell viability of cells that can be obtained. The aim of this preliminary study was to demonstrate the efficacy of an innovative medical device called Rigeneracons® (CE certified Class I) to provide autologous micro‐grafts immediately available to be used in the clinical practice. Moreover, Rigeneracons® is an instrument able to create micro‐grafts enriched of progenitors cells which maintain their regenerative and differentiation potential. We reported preliminary data about viability cell of samples derived from different kind of human tissues, such as periosteum, cardiac atrial appendage biopsy, and lateral rectus muscle of eyeball and disaggregated by Rigeneracons®. In all cases we observed that micro‐grafts obtained by Rigeneracons® displayed high cell viability. Furthermore, by cell characterization of periosteum samples, we also evidenced an high positivity to mesenchymal cell markers, suggesting an optimal regenerative potential. J. Cell. Physiol. 230: 2299–2303, 2015. © 2015 Wiley Periodicals, Inc.
Rachele Ciccocioppo, Giuseppina C. Cangemi, Peter Kruzliak, Alessandra Gallia, Elena Betti, Carla Badulli, Miryam Martinetti, Marila Cervio, Alessandro Pecci, Valeria Bozzi,et al.
Springer Science and Business Media LLC
Abstract Introduction Crohn’s disease (CD) is a disabling chronic enteropathy sustained by a harmful T-cell response toward antigens of the gut microbiota in genetically susceptible subjects. Growing evidence highlights the safety and possible efficacy of mesenchymal stem cells (MSCs) as a new therapeutic tool for this condition. Therefore, we aimed to investigate the effects of bone marrow-derived MSCs on pathogenic T cells with a view to clinical application. Methods T-cell lines from both inflamed and non-inflamed colonic mucosal specimens of CD patients and from healthy mucosa of control subjects were grown with the antigen muramyl-dipeptide in the absence or presence of donors’ MSCs. The MSC effects were evaluated in terms of T-cell viability, apoptotic rate, proliferative response, immunophenotype, and cytokine profile. The role of the indoleamine 2,3-dioxygenase (IDO) was established by adding a specific inhibitor, the 1-methyl-DL-tryptophan, and by using MSCs transfected with the small interfering RNA (siRNA) targeting IDO. The relevance of cell-cell contact was evaluated by applying transwell membranes. Results A significant reduction in both cell viability and proliferative response to muramyl-dipeptide, with simultaneous increase in the apoptotic rate, was found in T cells from both inflamed and non-inflamed CD mucosa when co-cultured with MSCs and was reverted by inhibiting IDO activity and expression. A reduction of the activated CD4+CD25+ subset and increase of the CD3+CD69+ population were also observed when T-cell lines from CD mucosa were co-cultured with MSCs. In parallel, an inhibitory effect was evident on the expression of the pro-inflammatory cytokines tumor necrosis factor-α, interferon-γ, interleukin-17A and -21, whereas that of the transforming growth factor-β and interleukin-6 were increased, and production of the tolerogenic molecule soluble HLA-G was high. These latter effects were almost completely eliminated by blocking the IDO, whose activity was upregulated in MSCs co-cultured with CD T cells. The use of a semipermeable membrane partially inhibited the MSC immunosuppressive effects. Finally, hardly any effects of MSCs were observed when T cells obtained from control subjects were used. Conclusion MSCs exert potent immunomodulant effects on antigen-specific T cells in CD through a complex paracrine and cell-cell contact-mediated action, which may be exploited for widespread therapeutic use.