Isolation protocols and comparative characterization of human mesenchymal stem cells derived from amnion and umbilical cord for regenerative medicine applications: a monocentric study

Introduction: The umbilical cord and the amniotic membrane are a precious source of human mesenchymal stem cells (hMSCs), even though they are often discarded after the delivery. Due to their immunomodulatory and antiinflammatory properties, hMSCs could be part of relevant strategies in the field of regenerative medicine. Additionally, they can be obtained from these tissues via a non-invasive and cost-effective process, overcoming ethical controversies. This study aims to propose protocol refinement to obtain hMSCs from the amnion and umbilical cord of healthy donors (utilizing limited and defined laboratory resources), and to compare and characterize these cells, thereby enabling future research on their properties. Methods: 30 women from “Paolo Giaccone” University Hospital of Palermo (Italy) were enrolled, according to the inclusion and exclusion criteria approved by the local Ethics Committee. A sample of umbilical cord and amnion was obtained from every patient and processed via an enzymatic or mechanical method. After refining the isolation protocol, hMSCs were characterized using flow cytometry, RT-qPCR, inducing a trilinear differentiation, and testing the formation of spheroids. Results: This research shows reliable and practical methods to isolate hMSCs from birth tissues, validating them with extensive cell characterization. No direct association was observed between mothers' age and newborns’ sex and the success rate in establishing hMSCs primary cultures, while a possible association between neonatal weight and the successful establishment of umbilical cord-derived cultures was found. Moreover, a difference in the adipogenic potential of the two hMSCs sources was highlighted. Discussion: hMSCs have a relevant role in biomedicine, along with their derivatives, for their promising regenerative properties: this study aims to explore new insights to promote further research in this field.