Generation of a neural stem progenitor cell lines (NPSCs), transfected with Green Fluorescent Protein (GFP) for in vivo monitoring

Abstract

[ES] Las células madre pluripotentes inducidas humanas (ihPSC) son consideradas universalmente una alternativa ética a las células madre embrionarias humanas. Al considerar la simplicidad de generar ihPSC y sus papeles previstos en la medicina regenerativa de crear células específicas de pacientes para su diferenciación en células adecuadas para la terapia celular, estas células suponen el candidato celular más prometedor para futuras terapias de sustitución celular incluso lesiones medulares. El objetivo de este proyecto es generar una línea estable de ihPSC transfectándola con el gen de la proteína fluorescente verde para su monitorización in vivo. Esta línea se caracterizará después de varios pases en el cultivo celular por los ensayos de inmunocitoquimica y aplicando el método de PCR en tiempo real para determinar si la transfeccion es estable y que no afecta la pluripotencia y la capacidad de diferenciación hacia progenitores neurales.

[EN] The presence of stem cells in the adult spinal cord (neural progenitor or epSPCs) entails an important progress in spinal cord injury treatment. This progress is not only related to cell therapy but also to enhance the endogenous regenerative potential of the nervous system. In the last few decades, the attention has been focused on studying the behavior of this stem cells or epSPCs in homeostasis or after the injury. Pharmacological studies, silencing with RNAi and overexpressing analysis with expression vectors in mammals are currently an excellent tool in order to increase or decrease cellular signaling pathways which upgrade this endogenous regeneration potential. Neural progenitors or epSPCs have an important role on cell therapy. Those stem cells help endogenous mechanism related to damage repairing and neural protection after spinal cord injury. It is, indeed, needed to insert a stable fluorescent marker, GFP for example, to these cells in order to study and monitor in vivo differentiated stem cells when transplanted. In first place, to observe the migration from the injection point to the lesion area and, in second place to check if cells incorporate themselves to the tissue structure in a natural way or if they form undesirable structures.