Abstract
Mesenchymal stromal cells (MSC) are used in cell therapy, but results depend on the unknown quality of cell populations. Extended culture time of MSC increases their senescent levels, leading to a critical loss of cell fitness. Here, we tested the suitability of MSC-sorting based on their FACS autofluorescence profile, for a rapid and non-invasive method of senescent cell elimination. Cells were classified in low- (LA) and high- (HA) autofluorescence groups, and results compared to the original MSC population (control). Three days after sorting, cells were screened by replicative senescence markers (cell volume, SA-β-Gal assay and gene/protein expression) and MSC differentiation assays. The transcriptional profiles of sorted MSC were also analyzed by RNA‐Seq. Compared to control, LA cells had 10% lower cell volume and autofluorescence, and 50% less SA-β-Gal + cells. Instead, HA cells had 20% higher cell volume and autofluorescence, and 120% more SA-β-Gal + cells. No changes in replicative senescence and differentiation potentials were observed between all groups. However, 68 genes (16 related to senescence) were significantly differentially expressed (DEG) between LA and other groups. Biological network of DEG identified CXCL12 as topological bottleneck. In summary, MSC sorting may have practical clinical implications to enhance the results of MSC-based therapies.
Highlights
Mesenchymal stromal cells (MSC) are used in cell therapy, but results depend on the unknown quality of cell populations
In comparison to the control group, average cell volume and autofluorescence were reduced in low average autofluorescence (LA) cells (10%), and increased in higher autofluorescence (HA) cells (30% and 20% respectively)
Based on system biology analysis, we identified the interaction network formed by the 155 differentially expressed genes (DEG) from the comparison between LA and HA cells (Fig. 7a) and the genes CXCL12, VCAM1 and LOX2 were recognized as a bottlenecks (Fig. 7b)
Summary
Mesenchymal stromal cells (MSC) are used in cell therapy, but results depend on the unknown quality of cell populations. The effectiveness and reliability of MSC in tissue regeneration may decrease due to age-related changes, both in vivo and in vitro, leading to a potential reduction in the MSC useful for cell-based therapy. This is relevant, in particular, for the medical use of autologous stem cell transplantation in elderly patients. A large number of cells are required, resulting from an extensive ex vivo cell expansion, and possibly leading to the accumulation of several senescent MSC phenotypes in culture. Our research has shown that MSC autofluorescence is directly related to cellular senescence measurements[17]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
