Growth and Differentiation Properties of Mesenchymal Stromal Cell Populations Derived from Whole Human Umbilical Cord

Abstract

Up to 2.8 × 10(7) fibroblast-like cells displaying an abundant presence of mesenchymal stem cell (MSC) markers CD73, CD90, CD105 and a low level of HLA-I expression can be isolated from one whole human umbilical cord (UC) using a simple and highly reproducible explant culture approach. Cells derived from whole UC, similar to cells collected from separate compartments of UC, display a distinct chondrogenic and adipogenic potential. Therefore they are potential candidates for cartilage and adipose tissue engineering. Cell differentiation along the osteogenic pathway is, however, less efficient, even after the addition of 1.25-dihydroxyvitamin D3, a potent osteoinductive substance. Isolated cells are highly proliferative, tolerate cryopreservation with an average survival rate of about 75% and after thawing can be propagated further, at least over 20 population doublings before their proliferative activity begins to decline. More importantly, they synthesize numerous trophic factors including neurotrophins and factors which facilitate angiogenesis and hematopoiesis. In conclusion, cells isolated from whole UC satisfies all requirements essential for the generation of stem cell banks containing permanently available cell material for applications in the field of regenerative medicine. Nevertheless, further studies are needed to improve and adjust the methods which are already employed for adult MSC expansion and differentiation to specific properties and requirements of the primitive stem cells collected from UC. So, our data verify that the choice of individual parameters for cell propagation, such as duration of cell expansion and cell seeding density, has a substantial impact on the quality of UC-derived cell populations.

Transplantation of umbilical cord–derived mesenchymal stem cells as a novel strategy to protect the central nervous system: technical aspects, preclinical studies, and clinical perspectives

“…Recently, a new method has been proposed for the isolation of MSCs from Wharton's jelly by the direct delivery of enzymatic solutions into the tissue, similar to the method used for the enzymatic digestion of the pancreas for Langerhans islet isolation (50). To avoid the disadvantages of enzymatic digestion, which has been shown to potentially alter cell proliferation and function, some groups have developed UC explant culture approaches that have been shown to be simple, reproducible, and efficient (63)(64)(65)(66). However, considering the heterogeneity of the processes used for cell isolation and expansion, and in view of their use for clinical applications, some groups have tried to develop optimized and standardized methods based on the use of xeno-and serum-free culture media (52,67,68) or by taking advantage of the plastic-adherence properties of MSCs that allow cell expansion without UC dissection or enzymatic digestion (69).…”

PDGFRβ Expression and Function in Fibroblasts Derived from Pluripotent Cells is Linked to DNA Demethylation

“…In light of these PDGFRb-mediated cellular functions, elucidating how expression of this receptor is regulated during development of mesenchymal cell types is of potential therapeutic value in understanding the mechanisms involved in tissue regeneration (Majore et al, 2011). However, as a result of numerous shortcomings of cells currently used for tissue repair strategies, deriving mesenchymal cells that manifest PDGFRbrelated functions from novel sources, such as induced pluripotent stem cells (iPSCs) and human embryonic stem cells (ESCs), might provide a renewable source of therapeutic cells for such regeneration strategies (Barberi et al, 2005;Riazi et al, 2009).…”

Isolation of Mesenchymal Stem Cells from Wharton’s Jelly in Comparison with Bone Marrow and Their Endothelial Differentiation

“…Failure was due to contamination and use of FBS. The explant method that we used was in agreement with Majore et al, [19]. This procedure is simple, reproducible and yields immunophenotypically homogenous cell population without enzymatic digestion of UC tissue.…”