Multi-Parameter Analysis of Biobanked Human Bone Marrow Stromal Cells Shows Little Influence for Donor Age and Mild Comorbidities on Phenotypic and Functional Properties

Abstract

Heterogeneous populations of human bone marrow-derived stromal cells (BMSC) are among the most frequently tested cellular therapeutics for treating degenerative and immune disorders, which occur predominantly in the aging population. Currently, it is unclear whether advanced donor age and commonly associated comorbidities affect the properties of ex vivo-expanded BMSCs. Thus, we stratified cells from adult and elderly donors from our biobank (n = 10 and n = 13, mean age 38 and 72 years, respectively) and compared their phenotypic and functional performance, using multiple assays typically employed as minimal criteria for defining multipotent mesenchymal stromal cells (MSCs). We found that BMSCs from both cohorts meet the standard criteria for MSC, exhibiting similar morphology, growth kinetics, gene expression profiles, and pro-angiogenic and immunosuppressive potential and the capacity to differentiate toward adipogenic, chondrogenic, and osteogenic lineages. We found no substantial differences between cells from the adult and elderly cohorts. As positive controls, we studied the impact of in vitro aging and inflammatory cytokine stimulation. Both conditions clearly affected the cellular properties, independent of donor age. We conclude that in vitro aging rather than in vivo donor aging influences BMSC characteristics.

Human Mesenchymal Stromal Cell (MSC) Characteristics Vary Among Laboratories When Manufactured From the Same Source Material: A Report by the Cellular Therapy Team of the Biomedical Excellence for Safer Transfusion (BEST) Collaborative

“…While the sample size of the study was not large enough to assess the nature of these differences, the analysis showed that the interlaboratory differences made a greater contribution to MSC variability than did differences among BM donors. This aligns with a recent study reporting that variable in vitro expansion strategies (i.e., passaging or "in vitro aging") have a stronger impact on MSC molecular phenotype than donor age (Andrzejewska et al, 2019).…”

Perirenal Adipose Tissue Displays an Age-Dependent Inflammatory Signature Associated With Early Graft Dysfunction of Marginal Kidney Transplants

“…While the quantitative distribution of PRAT-SVF endothelial cells was comparable in ECD and non-ECD donors, we observed a high inter-individual variability in PRAT-SVF angiogenic activity, that did not reach significance between the ECD and non-ECD groups analyzed in the present study. Such heterogeneity among donors has already been reported for mesenchymal stem cells (26). However, in line with this result, the study by Aird et al did not evidence major changes in the angiogenic potential of AT-SVF with aging except a delayed phase of neovessels maturation in vivo (46).…”

“…We and others have reported that the phenotypic and angiogenic activity of ECFC can be altered in deceased contexts associated with cardiovascular risk factors or genetic or epigenetic determinants (23)(24)(25). Furthermore, it was recently demonstrated that donor age and comorbidities can alter the angiogenic and paracrine immunosuppressive properties of human bone marrow-derived stromal cells (BM-SC) obtained after in vitro cell culture expansion (26). It is thus likely that the ECD microenvironment can also alter the vascular potential of the various types of PRAT SVF-resident cells.…”

Metal‐Specific Biomaterial Accumulation in Human Peri‐Implant Bone and Bone Marrow

“…MSC isolation and characterization was performed as described previously. [72] OBs were isolated from diced cancellous bone and characterized as described previously by others. [73] The cell culture medium for MSCs and OBs was Dulbecco's modified Eagle's medium with 10% fetal calf serum, 5 mm l-alanyl-lglutamine and 100 U mL −1 penicillin plus 100 µg mL −1 streptomycin.…”