Carfora, Antonia (2018) L'eterogeneità dei fibroblasti dermici come fattore in grado di condizionare l'efficacia della riprogrammazione cellulare. Dermal fibroblasts heterogeneity as a major factor influencing the efficiency of cell reprogramming. [Tesi di dottorato]

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Fibroblasts are a heterogeneous cell population as testified by the absence of specific markers and by their documented retention of positional memory. Fibroblasts have recently gained much attention as they are the adult somatic cell most commonly used for reprogramming to induced pluripotent stem cells (iPSCs). iPSCs are somatic cells genetically reprogrammed to a pluripotent state that makes them resemble embryonic stem cells (ESCs). iPSCs are a very promising tool for regenerative medicine because of their autologous origin and their ability to differentiate towards cell of all three germ layers. However, the iPSCs reprogramming technology is still affected by variable and low efficiency. We hypothesize that the heterogeneity of fibroblasts may play a role in determining cell reprogramming. To test our hypothesis, we isolated and cultured human dermal fibroblasts (HDFs) from five different anatomic sites (abdomen, breast, neck, thigh and arm) and performed a comparative study analyzing their morphology, proliferation, apoptosis and ability to migrate. Further, we evaluated whether the expression of mesenchymal and epithelial markers, the ability to differentiate and release growth factors in culture medium varied among fibroblasts from different anatomic sites. Our results revealed that, although fibroblasts from different anatomic sites had similar morphology, all other features varied according to the region from which they were isolated. Furthermore, we observed similarity between fibroblasts and mesenchymal stem cells (MSCs) that also varied among HDFs populations. A low proliferation index, the release of specific growth factors, the expression of mesenchymal markers and the promptness to differentiate towards mesodermal lineages, might improve the efficiency of reprogramming. On these bases, abdomen dermal fibroblasts might represent the ideal candidates for reprogramming; since they have a low proliferation index, express high amount of mesenchymal markers, differentiate more precociously and release growth factors that improve the efficiency of reprogramming. Further the high expression of mesenchymal markers and their ability to differentiate more precociously suggest that fibroblasts isolated from abdomen skin could represent a less differentiated phenotype, more similar to MSCs than the other population of fibroblasts and probably easier to reprogram. Therefore, our data offer novel perspectives in the field of cell reprogramming, suggesting that the choice of the dermal population of fibroblasts might influence the efficiency of reprogramming itself.

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