Isolation of mineralizing Nestin+ Nkx6.1+ vascular muscular cells from the adult human spinal cord

Background: The adult central nervous system (CNS) contains different populations of immature cells that could possibly be used to repair brain and spinal cord lesions. The diversity and the properties of these cells in the human adult CNS remain to be fully explored. We previously isolated Nestin(+) Sox2(+) neural multipotential cells from the adult human spinal cord using the neurosphere method (i.e. non adherent conditions and defined medium). -- Results: Here we report the isolation and long term propagation of another population of Nestin(+) cells from this tissue using adherent culture conditions and serum. QPCR and immunofluorescence indicated that these cells had mesenchymal features as evidenced by the expression of Snai2 and Twist1 and lack of expression of neural markers such as Sox2, Olig2 or GFAP. Indeed, these cells expressed markers typical of smooth muscle vascular cells such as Calponin, Caldesmone and Acta2 (Smooth muscle actin). These cells could not differentiate into chondrocytes, adipocytes, neuronal and glial cells, however they readily mineralized when placed in osteogenic conditions. Further characterization allowed us to identify the Nkx6.1 transcription factor as a marker for these cells. Nkx6.1 was expressed in vivo by CNS vascular muscular cells located in the parenchyma and the meninges. -- Conclusion: Smooth muscle cells expressing Nestin and Nkx6.1 is the main cell population derived from culturing human spinal cord cells in adherent conditions with serum. Mineralization of these cells in vitro could represent a valuable model for studying calcifications of CNS vessels which are observed in pathological situations or as part of the normal aging. In addition, long term propagation of these cells will allow the study of their interaction with other CNS cells and their implication in scar formation during spinal cord injury.

ENEKuS – a key model for managing the transformation of the normalisation of the Basque language in the workplace

The aim of this study is to develop a reference model for intervention in the language processes applied to the transformation of language normalisation within organisations of a socio-economic nature. It is based on the case study of an experience carried out over10 years within a trades’ union confederation, and has pursued a strategy of a basically qualitative research carried out in three stages: 1) undertaking field work through application of action-research methodology, 2) reconstructing experiences following processes of systematisation and conceptualisation of the systematised data, applying methodologies for the Systematisation of Experiences and Grounded Theory, and 3) formulating a model for intervention, applying the Systems Approach methodology. Finally, we identified nine key ideas that make up the conceptual framework for the ENEKuS reference model, which is structured in nine ‘action points', each having an operating sub-model applicable in practice.