Human adipose-derived mesenchymal stromal cells from face and abdomen undergo replicative senescence and loss of genetic integrity after long-term culture

Exp Cell Res. 2021 Sep 1;406(1):112740. doi: 10.1016/j.yexcr.2021.112740. Epub 2021 Jul 22.

Abstract

Body fat depots are heterogeneous concerning their embryonic origin, structure, exposure to environmental stressors, and availability. Thus, investigating adipose-derived mesenchymal stromal cells (ASCs) from different sources is essential to standardization for future therapies. In vitro amplification is also critical because it may predispose cell senescence and mutations, reducing regenerative properties and safety. Here, we evaluated long-term culture of human facial ASCs (fASCs) and abdominal ASCs (aASCs) and showed that both met the criteria for MSCs characterization but presented differences in their immunophenotypic profile, and differentiation and clonogenic potentials. The abdominal tissue yielded more ASCs, and these had higher proliferative potential, but facial cells displayed fewer mitotic errors at higher passages. However, both cell types reduced clonal efficiency over time and entered replicative senescence around P12, as evaluated by progressive morphological alterations, reduced proliferative capacity, and SA-β-galactosidase expression. Loss of genetic integrity was detected by a higher proportion of cells showing nuclear alterations and γ-H2AX expression. Our findings indicate that the source of ASCs can substantially influence their phenotype and therefore should be carefully considered in future cell therapies, avoiding, however, long-term culture to ensure genetic stability.

Keywords: Cell therapy; Fat depots; H2AX; MSC; Mesenchymal stem cells; Neural crest.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abdomen
  • Adipocytes / cytology
  • Adipocytes / metabolism*
  • Adipose Tissue / cytology
  • Adipose Tissue / metabolism*
  • Adult
  • Biomarkers / metabolism
  • Cell Differentiation
  • Cell Lineage / genetics*
  • Cell Proliferation
  • Cellular Senescence
  • Chondrocytes / cytology
  • Chondrocytes / metabolism*
  • Clone Cells
  • Face
  • Female
  • Gene Expression
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism*
  • Middle Aged
  • Mitosis
  • Organ Specificity
  • Osteocytes / cytology
  • Osteocytes / metabolism*
  • Phenotype
  • Primary Cell Culture
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • Biomarkers
  • H2AX protein, human
  • Histones
  • beta-Galactosidase