Optimization of umbilical cord processing techniques and multiparametric immunophenotyping of umbilical cord Wharton's jelly mesenchymal stem cells for cell therapy applications

Overview & Synopsis

With around 136,000,000 births per year worldwide stem cells isolated from neonatal tissues such as the umbilical cord have many advantages in term of access and availability. The umbilical cord contains a connective tissue present in large quantities surrounding the blood vessels, the Wharton’s jelly, demonstrated as a very rich source of Mesenchymal Stem Cells (MSCs). Those multipotent stem cells isolated from the umbilical cord have shown self‐renewal and differentiation capability into a range of cells inherent potential for cell therapy applications not only as differentiated cells but also thanks to potent capabilities for immunomodulation and regulation of inflammation. In this work we designed process and tools allowing efficient use of umbilical cord samples and the isolation of standardized Wharton's Jelly tissue explants. We studied techniques improving MSC isolation from tissue explants and developed an in‐vitro culture protocol, free of Foetal Bovine Serum (FBS) and other animal sourced products and thus more likely to be used in clinical settings. We developed a multiparametric flow cytometry protocol dedicated to phenotyping the surface expression of 27 different antigens present on Wharton's Jelly MSCs and to study the effect of several in‐vitro culture conditions and events on their surface phenotype. The multiparametric protocol was used to compare surface expression of MSCs isolated from Wharton's jelly, bone marrow and skin harvested fibroblasts. Finally, clinical development of human WJ‐MSC in a bone tissue differentiation protocol was evaluated for cleft palate repair, demonstrating the useful application of this tissue source