Cell quality evaluation with gene expression analysis of spheroids (3D) and adherent (2D) adipose stem cells

Gene. 2021 Feb 5:768:145269. doi: 10.1016/j.gene.2020.145269. Epub 2020 Oct 24.

Abstract

Adipose stem cells (ASCs) represent a reliable source of stem cells with a widely demonstrated potential in regenerative medicine and tissue engineering applications. New recent insights suggest that three-dimensional (3D) models may closely mimic the native tissue properties; spheroids from adipose derived stem cells (SASCs) exhibit enhanced regenerative abilities compared with those of 2D models. Stem cell therapy success is determined by "cell-quality"; for this reason, the involvement of stress signals and cellular aging need to be further investigated. Here, we performed a comparative analysis of genes connected with stemness, aging, telomeric length and oxidative stress, in 3D and 2D primary cultures. The expression levels of stemness-related markers and anti-aging Sirtuin1 were significantly up-regulated (P < 0.001) in SASCs-3D while gene expression of aging-related p16INK4a was increased in ASCs-2D (P < 0.001). The 3D and 2D cultures also had a different gene expression profile for genes related to telomere maintenance (Shelterin complex, RNA Binding proteins and DNA repair genes) (P < 0.01 and P < 0.001) and oxidative stress (aldehyde dehydrogenase class1 and 3) (P < 0.05, P < 0.01 and P < 0.001) and presented a striking large variation in their cellular redox state. Based on our findings, we propose a "cell quality" model of SASCs, highlighting a precise molecular expression of several genes involved with stemness (SOX2, POU5F1 and NANOG), anti-aging (SIRT1), oxidative stress (ALDH3) and telomeres maintenance.

Keywords: ALDH family; Adipose stem cells; Aging; Shelterin complex; Spheroid; Telomere length.

MeSH terms

  • Adipocytes / cytology
  • Adipocytes / metabolism*
  • Adipose Tissue / cytology*
  • Adolescent
  • Adult
  • Aged
  • Aging / genetics
  • Cell Adhesion / genetics
  • Cell Culture Techniques*
  • Cell Survival / genetics
  • Cell- and Tissue-Based Therapy / methods
  • Cells, Cultured
  • Cyclin-Dependent Kinase Inhibitor p16 / metabolism
  • DNA Repair / genetics
  • Female
  • Humans
  • Male
  • Middle Aged
  • Oxidative Stress / genetics
  • RNA-Binding Proteins / metabolism
  • Sirtuins / metabolism
  • Spheroids, Cellular / cytology
  • Stem Cell Transplantation*
  • Stem Cells / cytology*
  • Telomere Homeostasis / genetics
  • Tissue Engineering / methods
  • Young Adult

Substances

  • CDKN2A protein, human
  • Cyclin-Dependent Kinase Inhibitor p16
  • RNA-Binding Proteins
  • Sirtuins