This project’s primary aim was to pinpoint the tissue and processes originating the Amniotic Fluid Stem Cells (AFSCs), mesenchymal cells that can be consistently isolated from the human amniotic fluid. Since their identification in 2007, the AFSCs have been ascribed to the mesenchymal stem cells lineage and proposed as a tool for autologous regenerative medicine. However, despite several achievements and breakthroughs since their discovery in 2007 (De Coppi et al., Nature Botechnology 2007), none of the therapeutic approaches based on these cells has yet entered the clinic. This lack of functional translation can be ascribed to the poor knowledge on the origin and identity of the AFSCs. Hence, the objective of this project was to understand the cell’s identity and the biological processes leading to the AFSCs presence in the amniotic fluid. This would help in developing targeted therapeutic interventions that employ these cells for the functions that they are naturally programmed to exert, ultimately leading to the development of novel foetal and perinatal regenerative medicine strategies based on these cells. Overall, for this project I studied the transcriptomic profile and protein expression of the AFSC - in comparison to similar mesenchymal cells, isolated ad hoc from a library of foetal tissues. By highlighting the analogies and differences between these cell types, I aimed at inferring the foetal origin of the AFSCs, first by tissue, then by cell type and finally by anatomical location. To develop this project, I combined three available transcriptomic technologies (bulk RNA sequencing, single cell RNA sequencing and spatial transcriptomics). Together with protein validation assays like immunofluorescence, FACS analysis and western blot, this helped demonstrating the validity of my research hypothesis and further support my findings.
