A Comparative Analysis of Multipotent Mesenchymal Stromal Cells derived from Different Sources, with a Focus on Neuroregenerative Potential

Abstract

Multipotent mesenchymal stromal cells (MSCs) can be considered an accessible therapeutic tool for regenerative medicine. Here, we compared the growth kinetics, immunophenotypic and immunomodulatory properties, gene expression and secretome profile of MSCs derived from human adult bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and Wharton's jelly (WJ-MSCs) cultured in clinically-relevant conditions, with the focus on the neuroregenerative potential. All the cell types were positive for CD10/CD29/CD44/CD73/CD90/CD105/HLA-ABC and negative for CD14/CD45/CD235a/CD271/HLA-DR/VEGFR2 markers, but they differed in the expression of CD34/CD133/CD146/SSEA-4/MSCA-1/CD271/HLA-DR markers. BM-MSCs displayed the highest immunomodulatory activity compared to AT- and WJ-MSCs. On the other hand, BM-MSCs secreted the lower content and had the lower gene expression of neurotrophic growth factors compared to other cell lines, which may be caused by the higher sensitivity of BM-MSCs to nutrient limitations. Despite the differences in growth factor secretion, the MSC secretome derived from all cell sources had a pronounced neurotrophic potential to stimulate the neurite outgrowth of DRG-neurons and reduce the cell death of neural stem/progenitor cells after H₂O₂ treatment. Overall, our study provides important information for the transfer of basic MSC research towards clinical-grade manufacturing and therapeutic applications.

Figure 1

Immunophenotype of MSCs derived from different sources. Data are expressed as mean ± SEM, N = 7 (*p < 0.05; **p < 0.01; ***p < 0.001). One-way ANOVA with Student–Newman–Keuls post hoc pair-to-pair test.

Figure 2

Growth kinetics of MSCs, derived from different sources, assessed by cPD (A) and PDT (B). Data are expressed as mean ± SEM (N = 7 for BM-MSCs, N = 9 for AT-MSCs and N = 15 for WJ-MSCs). (*p < 0.05; **p < 0.01; ***p < 0.001). A - One-way ANOVA with Student–Newman–Keuls post hoc pair-to-pair test; B - One-way ANOVA with Dunn’s post hoc pair-to-pair test.

Figure 3

Immunomodulatory properties of MSCs derived from different sources in contact (A) or transwell (B) co-culture with PBMC (mean ± SEM, N = 3 in duplicates). *p < 0.05 compared to other sources; ^p < 0.05 compared to PBMC alone group. Mann-Whitney nonparametric test.

Figure 4

The secretome profile of MSCs of different tissue origin in serum starvation (PL-free) conditions (mean ± SEM, N = 3 in duplicates). *p < 0.05, ***p < 0.001. One-way ANOVA with Student–Newman–Keuls post hoc pair-to-pair test.

Figure 5

The effect of MSC-derived CM on the recovery of neural stem cells (SPC-01) after H₂O₂ treatment (mean ± SEM, N = 3 in duplicates). *p < 0.05 - values are significantly different compared to control. Mann-Whitney nonparametric test.

Figure 6

The effect of MSC-derived CM on neurite growth of rat DRG neurons: (A) DRG neurons cultured in the presence of conditioned or control medium (beta3 tubulin/DAPI staining, magn. ×200); (B) the fold change in mean area (mean ± SEM, N = 3 in duplicates) of neurite growth related to control (α-MEM). *p < 0.05 values are significantly higher compared to control (α-MEM). Mann-Whitney nonparametric test.

Figure 7

The effect of serum starvation (PL-free) conditions on metabolic activity of MSCs derived from different tissues (mean ± SEM, N = 3 in duplicates). *p < 0.05 values are significantly different compared to CCM group. Paired Student’s t-test.

Figure 8

Relative gene expression of ex vivo expanded MSCs derived from different tissues cultured in CCM or serum starvation (PL-free) conditions (mean ± SEM, N = 3 in duplicates for PL-free groups, N = 7 in duplicates for CCM groups). *p < 0.05; **p < 0.01; ***p < 0.001). One-way ANOVA.