Human Wharton’s jelly-derived mesenchymal stromal cells promote bone formation in immunodeficient mice when administered into a bone microenvironment

Bone microenvironment; Bone regeneration; Multipotent mesenchymal stromal cellsMicroambient ossi; Regeneració òssia; Cèl·lules estromals mesenquimals multipotentsMicroambiente óseo; Regeneración ósea; Células estromales mesenquimales multipotentesBackground Wharton’s Jelly (WJ) Mesenchymal Stromal Cells (MSC) have emerged as an attractive allogeneic therapy for a number of indications, except for bone-related conditions requiring new tissue formation. This may be explained by the apparent recalcitrance of MSC,WJ to differentiate into the osteogenic lineage in vitro, as opposed to permissive bone marrow (BM)-derived MSCs (MSC,BM) that readily commit to bone cells. Consequently, the actual osteogenic in vivo capacity of MSC,WJ is under discussion. Methods We investigated how physiological bone environments affect the osteogenic commitment of recalcitrant MSCs in vitro and in vivo. To this end, MSC of BM and WJ origin were co-cultured and induced for synchronous osteogenic differentiation in vitro using transwells. For in vivo experiments, immunodeficient mice were injected intratibially with a single dose of human MSC and bone formation was evaluated after six weeks. Results Co-culture of MSC,BM and MSC,WJ resulted in efficient osteogenesis in both cell types after three weeks. However, MSC,WJ failed to commit to bone cells in the absence of MSC,BM’s osteogenic stimuli. In vivo studies showed successful bone formation within the medullar cavity of tibias in 62.5% of mice treated with MSC, WJ. By contrast, new formed trabeculae were only observed in 25% of MSC,BM-treated mice. Immunohistochemical staining of human COXIV revealed the persistence of the infused cells at the site of injection. Additionally, cells of human origin were also identified in the brain, heart, spleen, kidney and gonads in some animals treated with engineered MSC,WJ (eMSC,WJ). Importantly, no macroscopic histopathological alterations, ectopic bone formation or any other adverse events were detected in MSC-treated mice. Conclusions Our findings demonstrate that in physiological bone microenvironment, osteogenic commitment of MSC,WJ is comparable to that of MSC,BM, and support the use of off-the-shelf allogeneic MSC,WJ products in bone repair and bone regeneration applications.This work was funded by CRG’s Emergent Translational Research Project Grant. Research in JV's laboratory is developed in the context of Red Española de Terapias Avanzadas (TERAV, expedient no. RD21/0017/0022) funded by Instituto de Salud Carlos III (ISCIII) in the context of NextGenerationEU's Recovery, Transformation and Resilience Plan, and has been recognised as a Consolidated Research Group by Generalitat de Catalunya (2021-SGR-00877). The disclosed funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Los niveles epicardiales de les Coves del Fem (Ulldemolins, Tarragona)

Les Coves del Fem (Ulldemolins) es una gran cavidad situada en el macizo del Montsant, a 490 m snm, en el interior de Tarragona. Los trabajos iniciados el año 2013 han permitido documentar una amplia estratigrafía que abarca desde el Mesolítico Reciente hasta el Neolítico Postcardial. Los resultados de las intervenciones arqueológicas ponen de relieve el interés de este yacimiento por lo se refiere a la transición entre los últimos cazadores recolectores y los primeros grupos de agricultores y ganaderos en el norestes peninsular. En el marco de un nuevo proyecto de investigación se ha llevado a cabo una nueva intervención arqueológica en la parte central del abrigo, en un área que ocupa 9 m2. La zona intervenida colinda con los perfiles realizados en las campañas de 2013 y 2015 sobre un corte resultante de intervenciones clandestinas previas. La intención es obtener, en un futuro próximo, una sección transversal completa desde este punto hasta el fondo de la cueva. La excavación en extensión de esta área ha hecho patente la complejidad estratigráfica del yacimiento donde las unidades se presentan de forma muy heterogénea con sucesiones de niveles de arenas de fracción muy fina depositadas por la acción de las crecidas del río Montsant y por sedimentación eólica. Aunque el área intervenida no es muy amplia se han documentado un número relevante de estructuras como fosas, elementos de sostenimiento y estructuras de combustión de diversa tipología y función, algunas de las cuales se organizan en relación a una gran estructura compleja que, pese a que debería confirmarse en futuras intervenciones, podría interpretarse como una base de una cabaña. Los niveles excavados hasta la fecha corresponden a las ocupaciones epicardiales documentadas en las intervenciones anteriores, las cuales han proporcionado un abundante y variado conjunto material. Cabe destacar la industria lítica tallada en sílex, que abarca restos representativos de todas las fases del proceso de producción, la industria ósea, los elementos cerámicos, los restos antracológicos y carpológicos, así como restos de madera no carbonizada o semicarbonizada en buen estado de conservación.Peer reviewe

Efficient Generation of Germ Line Transmitting Chimeras from C57BL/6N ES Cells by Aggregation with Outbred Host Embryos

Genetically modified mouse strains derived from embryonic stem (ES) cells have become essential tools for functional genomics and biomedical research. Large scale mutagenesis projects are producing libraries of mutant C57BL/6 (B6) ES cells to enable the functional annotation of every gene of the mouse genome. To realize the utility of these resources, efficient and accessible methods of generating mutant mice from these ES cells are necessary. Here, we describe a combination of ICR morula aggregation and a chemically-defined culture medium with widely available and accessible components for the high efficiency generation of germline transmitting chimeras from C57BL/6N ES cells. Together these methods will ease the access of the broader biomedical research community to the publicly available B6 ES cell resources

Pleiotropy of Glycogen Synthase Kinase-3 Inhibition by CHIR99021 Promotes Self-Renewal of Embryonic Stem Cells from Refractory Mouse Strains

Background: Inhibition of glycogen synthase kinase-3 (GSK-3) improves the efficiency of embryonic stem (ES) cell derivation from various strains of mice and rats, as well as dramatically promotes ES cell self-renewal potential. b-catenin has been reported to be involved in the maintenance of self-renewal of ES cells through TCF dependent and independent pathway. But the intrinsic difference between ES cell lines from different species and strains has not been characterized. Here, we dissect the mechanism of GSK-3 inhibition by CHIR99021 in mouse ES cells from refractory mouse strains. Methodology/Principal Findings: We found that CHIR99021, a GSK-3 specific inhibitor, promotes self-renewal of ES cells from recalcitrant C57BL/6 (B6) and BALB/c mouse strains through stabilization of b-catenin and c-Myc protein levels. Stabilized b-catenin promoted ES self-renewal through two mechanisms. First, b-catenin translocated into the nucleus to maintain stem cell pluripotency in a lymphoid-enhancing factor/T-cell factor–independent manner. Second, b-catenin binds plasma membrane-localized E-cadherin, which ensures a compact, spherical morphology, a hallmark of ES cells. Further, elevated c-Myc protein levels did not contribute significantly to CH-mediated ES cell self-renewal. Instead, the role of c-Myc is dependent on its transformation activity and can be replaced by N-Myc but not L-Myc. b-catenin and c-Myc have similar effects on ES cells derived from both B6 and BALB/c mice. Conclusions/Significance: Our data demonstrated that GSK-3 inhibition by CH promotes self-renewal of mouse ES cell

Isolation of Oct4-Expressing Extraembryonic Endoderm Precursor Cell Lines

BACKGROUND:The extraembryonic endoderm (ExEn) defines the yolk sac, a set of membranes that provide essential support for mammalian embryos. Recent findings suggest that the committed ExEn precursor is present already in the embryonic Inner Cell Mass (ICM) as a group of cells that intermingles with the closely related epiblast precursor. All ICM cells contain Oct4, a key transcription factor that is first expressed at the morula stage. In vitro, the epiblast precursor is most closely represented by the well-characterized embryonic stem (ES) cell lines that maintain the expression of Oct4, but analogous ExEn precursor cell lines are not known and it is unclear if they would express Oct4. METHODOLOGY/PRINCIPAL FINDINGS:Here we report the isolation and characterization of permanently proliferating Oct4-expressing rat cell lines ("XEN-P cell lines"), which closely resemble the ExEn precursor. We isolated the XEN-P cell lines from blastocysts and characterized them by plating and gene expression assays as well as by injection into embryos. Like ES cells, the XEN-P cells express Oct4 and SSEA1 at high levels and their growth is stimulated by leukemia inhibitory factor, but instead of the epiblast determinant Nanog, they express the ExEn determinants Gata6 and Gata4. Further, they lack markers characteristic of the more differentiated primitive/visceral and parietal ExEn stages, but exclusively differentiate into these stages in vitro and contribute to them in vivo. CONCLUSIONS/SIGNIFICANCE:Our findings (i) suggest strongly that the ExEn precursor is a self-renewable entity, (ii) indicate that active Oct4 gene expression (transcription plus translation) is part of its molecular identity, and (iii) provide an in vitro model of early ExEn differentiation

The Liberation of Embryonic Stem Cells

Mouse embryonic stem (ES) cells are defined by their capacity to self-renew and their ability to differentiate into all adult tissues including the germ line. Along with efficient clonal propagation, these properties have made them an unparalleled tool for manipulation of the mouse genome. Traditionally, mouse ES (mES) cells have been isolated and cultured in complex, poorly defined conditions that only permit efficient derivation from the 129 mouse strain; genuine ES cells have not been isolated from another species in these conditions. Recently, use of small molecule inhibitors of glycogen synthase kinase 3 (Gsk3) and the Fgf-MAPK signaling cascade has permitted efficient derivation of ES cells from all tested mouse strains. Subsequently, the first verified ES cells were established from a non-mouse species, Rattus norvegicus. Here, we summarize the advances in our understanding of the signaling pathways regulating mES cell self-renewal that led to the first derivation of rat ES cells and highlight the new opportunities presented for transgenic modeling on diverse genetic backgrounds. We also comment on the implications of this work for our understanding of pluripotent stem cells across mammalian species

Famílies botàniques de plantes medicinals

Facultat de Farmàcia, Universitat de Barcelona. Ensenyament: Grau de Farmàcia, Assignatura: Botànica Farmacèutica, Curs: 2013-2014, Coordinadors: Joan Simon, Cèsar Blanché i Maria Bosch.Els materials que aquí es presenten són els recull de 175 treballs d’una família botànica d’interès medicinal realitzats de manera individual. Els treballs han estat realitzat per la totalitat dels estudiants dels grups M-2 i M-3 de l’assignatura Botànica Farmacèutica durant els mesos d’abril i maig del curs 2013-14. Tots els treballs s’han dut a terme a través de la plataforma de GoogleDocs i han estat tutoritzats pel professor de l’assignatura i revisats i finalment co-avaluats entre els propis estudiants. L’objectiu principal de l’activitat ha estat fomentar l’aprenentatge autònom i col·laboratiu en Botànica farmacèutica

Les construccions en pedra i àrees d'activitat associades

[EN] The stone constructions characterize the most recent phase of occupation of the settlement. They appear as a paved surface that covers and amortizes the previous constructions made of wood. Several combustion areas and structures for food conservation have been documented. The distribution of the pavement and stone structures are unequal across the whole excavated area of the site. The pavements are made up of large, small and medium-sized travertine slabs. They display particular formal characteristics that make it very difficult to make generalizations. However, there are sorne common traits such as the presence around them and also inside them of holes for wooden poles. The hearths identified are characterized by travertine blocks, stoneware or burnt pebbles, ali in varying quantities, and charcoal, which is the remains of the firewood used. They are very numerous in Sector A, where about forty have been found of different morphologies. The pits, during this second phase o/ occupation, are slightly depressed constructions, of irregular shapes and both small and large in size, that are filled with large quantities of diverse archaeological materials such as charred seeds, animal bones, potsherds, quartz, flint, bone tools, pieces of ornaments and grinding instruments. Because of the material they contain, the pits have been interpreted as middens lor food waste and manufactured objects that were once considered useless.[CA] Les construccions realitzades en pedra caracteritzen la fase més recent d'ocupació del poblat. Aquestes construccions es presenten com paviments enllosats que cobreixen i amortitzen les construccions anteriors realitzades en fusta. Al llarg de la seva superticie s'evidencia la presència d'àrees de combustió i d'estructures dedicades a la conservació d'aliments. Aquesta situació es constata amb diferent intensitat en tota l'àrea excavada del jaciment. Els paviments identificats es construeixen a partir de lloses de travertí de dimensions dispars, arribant al seu supetiicie diferent continuïtat. Aquests paviments mostren entre ells característiques formals particulars que dificulten la realització de generalitzacions. No obstant això, entre aquestes s'observa la presència de forats de pals de fusta al seu voltant i també al seu interior. Les llars identificats es caracteritzen per contenir blocs de travertí, plaques de gres i còdols, normalment cremats i en quantitat variable, a més de carbons corresponents a les restes de combustible utilitzat. Les llars són molt nombrosos en el sector A, on s'han documentat més de quaranta, i presenten mariologías variables: plana, en forma de cubeta i de cubeta-forn. En aquesta fase d'ocupació més recent, les fosses s'assimilen construccions lleugerament deprimides, de contorn irregular i diferents dimensions. Habitualment es troben farcides de grans quantitats de restes arqueològiques diversos com llavors carbonitzades, ossos d'animals, fragments de ceràmica, restes lítiques, instrumental de mòlta, eines d'os i elements d'adorn personal. Pel seu contingut aquestes fosses han estat interpretades com a abocadors de residus alimentaris i de deixalles procedents de la manufactura de diferents tipus de béns i estris.[ES] Las construcciones realizadas en piedra caracterizan la fase más reciente de ocupación del poblado. Estas construcciones se presentan como pavimentos enlosados que cubren y amortizan las construcciones anteriores realizadas en madera. A lo largo de su superticie se evidencia la presencia de áreas de combustión y de estructuras dedicadas a la conservación de alimentos. Esta situación se constata con distinta intensidad en toda el área excavada del yacimiento. Los pavimentos identificados se construyen a partir de losas de travertino de dimensíones dispares, alcanzando su supetiicie distinta continuidad. Estos pavimentos muestran entre ellos características formales particulares que dificultan la realización de generalizaciones. Sin embargo, entre estas se observa la presencia de agujeros de postes de madera a su alrededor y también en su interior. Los hogares identificados se caracterizan por contener bloques de travertino, placas de arenisca y cantos rodados, normalmente quemados y en cantidad variable, además de carbones correspondientes a los restos de combustible utilizado. Los hogares son muy numerosos en el sector A, donde se han documentado más de cuarenta, y presentan mariologías variables: plana, en forma de cubeta y de cubeta-horno. En esta fase de ocupación más reciente, las fosas se asimilan a construcciones ligeramente deprimidas, de contorno irregular y distintas dimensiones. Habitualmente se hallan rellenas de grandes cantidades de restos arqueológicos diversos como semillas carbonizadas, huesos de animales, fragmentos de cerámica, restos líticos, instrumental de molienda, herramientas de hueso y elementos de adorno personal. Por su contenido dichas fosas han sido interpretadas como vertederos de residuos alimentarios y de desechos procedentes de la manufactura de distintos tipos de bienes y utensilios.Peer reviewe

The challenge of developing human 3D organoids into medicines

The capacity of organoids to generate complex 3D structures resembling organs is revolutionizing the fields of developmental and stem cell biology. We are currently establishing the foundations for translational applications of organoids such as drug screening, personalized medicine and launching the future of cell therapy using organoids. However, clinical translation of organoids into cell replacement therapies is halted due to (A) a few preclinical studies demonstrating their efficacy and (B) the lack of robust, reproducible, and scalable methods of production in compliance with current pharmaceutical standards. In this issue of Stem Cell Research & Therapy [ref], Dossena and collaborators present a validated bioprocess design for large-scale production of human pancreatic organoids from cadaveric tissue in accordance with current good manufacturing practice. The authors also propose a set of specifications of starting materials and critical quality attributes of final products that are of interest to other developments provided that this type of medicines are different than any other medicinal product due to their complex composition and living nature of the active ingredient. Although large-scale production of functional cells secreting insulin is still a challenge, the development of methods such as the one presented by Dossena and collaborators contributes to move toward clinical use of organoids in the treatment of type 1 diabetes and opens avenues for future clinical use of organoids in degenerative pathologies.Work in JV’s laboratory is supported by the Spanish Cell Therapy Network (TerCel, expedient no. RD16/0011/0028), awarded by Generalitat de Catalunya as Consolidated Research Group (ref. 2017-SGR-719). Project PI19/ 01788 is funded by Instituto de Salud Carlos III and co-funded by European Union (ERDF/ESF) - A way to build Europ