Maternal Serum, an Isolation and Expansion Tool for Umbilical Cord Matrix Mesenchymal Stromal Cells

Abstract

The umbilical cord offers a source of readily available mesenchymal stromal cells (MSCs) for use in research and ultimately therapeutic application. However, methods of isolating these cells vary between investigators, and no standard method has been adopted. The aims of this work were to (i) develop a methodology for the isolation of umbilical cord matrix cells without the use of enzymatic digestion or complicated dissection; (ii) investigate the use of pooled maternal serum (MS) as a media supplement; and (iii) demonstrate that the cells isolated were MSCs. We have demonstrated that incubating tissue explants of less than 2 mm3 in serum for an hour, followed by the gradual addition of serum containing culture medium can increase cell yield compared to incubation in serum containing culture medium alone. More importantly, our method demonstrated that the use of pooled serum from women >37 weeks pregnant (pooled MS) yields higher cell numbers than the use of fetal bovine serum or pooled umbilical cord serum. Irrespective of the type of serum used, the isolated cells were MSCs according to the minimal criteria set out by the Mesenchymal and Tissue Stem Cell Committee of the International Society for Cellular Therapy. In conclusion, MS has the potential to be used as an alternative to fetal bovine serum for isolation and expansion of umbilical cord MSCs for clinical purposes.