Isolation method and xeno-free culture conditions influence multipotent differentiation capacity of human Wharton’s jelly-derived mesenchymal stem cells

Abstract

IntroductionHuman Wharton’s jelly (WJ) has become a preferred source of mesenchymal stem cells (MSCs) whose clinical applications are limited by the use of adequate xeno-free (XF), in vitro manipulation conditions. Therefore, the objective of our study was to characterize WJ-derived MSCs (WJ-MSCs), isolated by different methods and cultured in a commercially available, MSC XF medium, not least of all by investigating their endothelial differentiation capacity.MethodsWJ explants and enzymatically dissociated WJ cells were cultured in a defined, XF medium for MSCs. Adherent cells at passages 2 and 5 were characterized as MSCs by flow cytometry, MTT, real-time quantitative reverse transcription PCR, and functional multipotent differentiation assays. The endothelial differentiation capacity of MSCs isolated and expanded until passage 2 in the MSC XF medium, and then subcultured for five passages in a commercially available endothelial growth medium (group A), was assessed over serial passages, as compared to adherent WJ-derived cells isolated and expanded for five consecutive passages in the endothelial medium (group B).ResultsThe MSC phenotype of WJ explant- and pellet-derived cells, isolated and expanded in the MSC XF medium, was proven based on the expression of CD44/CD73/CD90/CD105 surface markers and osteo-/adipo-/chondrogenic multipotent differentiation potential, which differed according to the isolation method and/or passage number. Upon exposure to endothelial differentiation cues, cells belonging to group A did not exhibit endothelial cell characteristics over serial passages; by contrast, WJ pellet-derived cells belonging to group B expressed endothelial characteristics at gene, protein and functional levels, potentially due to culture conditions favoring the isolation of other stem/progenitor cell types than MSCs, able to give rise to an endothelial progeny.ConclusionsThe use of defined, MSC XF media for isolation and expansion of human WJ-MSCs is a prerequisite for the establishment of their real endothelial differentiation capacity, as candidates for clinical therapy applications. Thus, the standardization of WJ-MSCs isolation and culture expansion techniques in defined, MSC XF media, for their accurate characterization, would be a priority in the stem cell research field.

Differentiation of Human Umbilical Cord Matrix Mesenchymal Stem Cells into Neural-Like Progenitor Cells and Maturation into an Oligodendroglial-Like Lineage

“…Nevertheless, we observed low adipogenic induction, a characteristic which has already been reported in the literature for UCM-MSCs, when compared with MSCs from other sources, such as the bone-marrow [53].…”

Integrated culture platform based on a human platelet lysate supplement for the isolation and scalable manufacturing of umbilical cord matrix-derived mesenchymal stem/stromal cells

“…However, foreseeing the therapeutic use of these cells, efforts are being made to avoid the use of FBS because of the high risk of contamination (virus, prion), potential to promote xenogeneic immune reactions (Bieback, ) and the ethical issues associated with the harvesting and collection of this animal‐derived product (Gstraunthaler, ). In this context, several groups, including the present authors, have demonstrated the ability to isolate and expand UCM MSCs in planar tissue culture systems using more defined serum‐/xenogeneic(xeno)‐free culture medium preparations (Corotchi et al , ; Simões et al , ; Swamynathan et al , ). Others have tested the use of human blood‐derived platelet lysate (HPL) products as culture medium supplements for this purpose.…”

Discarded Wharton jelly of the human umbilical cord: a viable source for mesenchymal stromal cells

“…The cardiovascular system has low regenerative potential, which makes the use of WJ-MSCs an ideal alternative in cardiovascular tissue repair with their immunomodulatory properties and self-regenerating capacity [26]. Interestingly, biologically active heart valve leaflets could be engineered using only cells from the human umbilical cord [27].…”