Influence of Donor Age and Passage Number on Angiogenic Activity in Human Adipose-Derived Stem Cell-Conditioned Media

Abstract

Introduction: Adipose-derived stem/stromal cells (ASCs) are considered a promising cell source for therapeutic angiogenesis because the cells can be prepared following a minimally invasive procedure and because they secrete a variety of angiogenic cytokines. In the present study, the influence of donor age and passage number on angiogenic activity of ASC-conditioned media (ASC-CM) was examined.Methods: Human ASCs (donor age, 5 months to 82 years; n = 10) were cultured, and ASC-CM were collected at passages 2, 4, and 6. The angiogenic activity of ASC-CM was evaluated with the tube formation assay using a system in which human umbilical vein endothelial cells (HUVECs) and fibroblasts were co-cultured. The concentrations of vascular endothelial growth factor-A (VEGF-A) and hepatocyte growth factor (HGF) in each ASC-CM were measured using an enzyme-linked immunosorbent assay. Results:A donor age over 60 years affected the proliferative capacity of ASCs at passage 4 and later. ASC-CM significantly enhanced HUVEC tube formation, and this response was not influenced by donor age. ASC-CM at passage 6 showed a lower tube formation capacity compared to ASC-CM at passage 4, although the capacity was still equivalent to the positive control (medium containing10 ng/mL VEGF-A). A donor age over 26 years affected VEGF-A but not HGF levels in ASC-CM, although we found no direct correlation between VEGF-A/HGF levels and the tube formation capacity. Conclusion:Our results demonstrate a passage number-dependent but a donor age-independent decline in the angiogenic activity of ASC-CM.

Secretome of Senescent Adipose-Derived Mesenchymal Stem Cells Negatively Regulates Angiogenesis

“…It is worth noting that the authors used simplified models close to the in vivo conditions. ASC-CM-induced HUVEC tube formation was evaluated capacity in vitro only [34]. Thus, based on our results and the data of experiments using ASCs from the donors of various ages, we can conclude that senescent ASCs have a lesser angiogenic activity versus "young" cells.…”

Quantitative Analysis of Factors Regulating Angiogenesis for Stem Cell Therapy

“…Still, it is thought that the secretion of cytokines from DFAT cells mainly promotes the tube formation of vascular endothelial cells. Human adipose-derived stem/stromal cells (ASCs) could differentiate into adipocyte, osteoblasts, chondrocytes, and smooth muscle-like cells [ 34 ]. The conditioned medium of ASCs (ASC-CM) enhanced HUVEC tube formation (Kurabo), and this response was not influenced by donor age.…”

“…The conditioned medium of ASCs (ASC-CM) enhanced HUVEC tube formation (Kurabo), and this response was not influenced by donor age. Considerable levels of VEGF-A and HGF were detected in ASC-CM measured with ELISA [ 34 ].…”