In the past years, biomedical research has been strongly oriented toward the design of tools and techniques aimed at providing personalized therapeutical approaches and tailored treatments. In this perspective, the possibility to study, in vitro, patient cells that would normally be inaccessible with conventional clinical investigation tools has become critically relevant. The introduction of reprogramming techniques to convert adult somatic cells (i.e. skin fibroblasts) into pluripotent cells earned Prof. Shin’ya Yamanaka the Nobel Prize and is currently considered one of the greatest achievement in biology of the last decades. Induced pluripotent stem (iPS) cells, by definition, can be differentiated virtually toward all cell types of the body, similarly to embryonic stem cells, but with the major advantages of bypassing ethical concerns and avoiding issues related to immune rejection for cell replacement therapies in humans. Of note, the possibility to generate platforms of cells to validate the efficacy of novel drugs or to elucidate pathological mechanisms has become increasingly important especially in the Neuroscience field, which for years has been limited by restricted access to living human neurons.
On this basis, the Laboratory of Cellular Engineering is oriented to the generation of iPS cells from human fibroblasts and to the differentiation of reprogrammed pluripotent cells into neural stem cells amenable to be terminally converted into mature neurons and glial cells. The generation of highly expandable neural stem cells has been demonstrated to be useful to guarantee a virtually unlimited access to neurons with diverse positional identity and allows us to investigate several pathological conditions including neurodevelopmental and neurodegenerative disorders. For this purposes the Laboratory of Cellular Engineering is fully equipped with state-of-the-art instrumentations to allow appropriate manipulation of human iPS cells and neural stem cells. Specific instrumentations include:
Laminar Flow Hood for cell manipulation under adequate sterile conditions.
Fluorescence Stereomicroscope (StereoDiscovery V8, Zeiss) with magnification power from 10X to 80X located under vertical laminar flow hood for iPS cell colony picking in sterility. The microscope is equipped with fotocamera and adapted workstation for image acquisition, post-processing and linear measurements.
Nucleofector 4D for cell transfection of reprogramming episomal vectors in suspension.
The Laboratory also takes advantage of the Imaging Facility at FPS for cell analysis and characterization. In particular, image acquisitions are routinely performed with Single-Photon Confocal Microscope (Olympus FV3000) and a Super-Resolution Microscope for detection of cellular ultrastructure.
The Cellular Engineering Laboratory is directed by Maria Teresa Dell’Anno