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Peer-reviewedCC BY-NCAdipocyte

Comparative analysis of select 3D human mesenchymal stromal cell-derived spheroid models for adipogenic differentiation.

  • Bianca Vezzani1
  • Elisa Tamburini1
  • Anna Terenziani1
  • Nadia Ilaria Oliverio1
  • Simonetta Croci2
  • Alessandro Bartolomucci1,3
  1. 1Laboratory of Biogerontology and Geroscience, Department of Medicine and Surgery, University of Parma, Parma, Italy.
  2. 2Department of Medicine and Surgery, University of Parma, Parma, Italy.
  3. 3Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, USA.
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Abstract

In vitro modelling of adipose tissue (AT) commonly employs 2D systems, which do not fully capture the complexity of AT, lacking cell-cell interactions and extracellular matrix (ECM) remodelling. Despite the innovations in 3D culture systems offering physiologically relevant AT platforms, the field lacks consensus and cross-protocol method comparison. We performed a comparative analysis of three scaffold-free spheroid-generation techniques: Ultra Low Attachment plates (ULA), Hanging Drops (HD), and Microwell Culture Plates (MCP), utilizing human AT-derived mesenchymal stromal cells. Each method produced viable spheroids of similar size and morphology within 48 hours. However, MCP demonstrated greater uniformity and a significant increase in mass density when compared to ULA and HD. Adipogenic media increased spheroid size in ULA and MCP, but not in HD spheroids. Notably, the diameter of ULA spheroids was 2- and 6-times larger compared to HD and MCP ones, respectively, regardless of adipogenic induction. Gene expression analysis indicated that all techniques led to PPARγ and ADIPOQ expression during adipogenesis. All methods supported adipocyte differentiation, as evidenced by lipid droplet accumulation, PLIN1, CD36, and HSL expression, and ECM rearrangement. These findings suggest that all the described techniques constitute a robust platform for producing adipogenesis-competent spheroids, while method-specific differences suggest an application-dependent choice.

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Cite

BibTeXRIS
BibTeX
@article{vezzani2026comparative,
  title = {{Comparative analysis of select 3D human mesenchymal stromal cell-derived spheroid models for adipogenic differentiation.}},
  author = {Bianca Vezzani and Elisa Tamburini and Anna Terenziani and Nadia Ilaria Oliverio and Simonetta Croci and Alessandro Bartolomucci},
  journal = {Adipocyte},
  year = {2026},
  month = oct,
  doi = {10.1080/21623945.2026.2736340},
  url = {https://europepmc.org/article/MED/42852735},
  note = {CC BY-NC},
}
RIS
TY  - JOUR
TI  - Comparative analysis of select 3D human mesenchymal stromal cell-derived spheroid models for adipogenic differentiation.
AU  - Bianca Vezzani
AU  - Elisa Tamburini
AU  - Anna Terenziani
AU  - Nadia Ilaria Oliverio
AU  - Simonetta Croci
AU  - Alessandro Bartolomucci
PY  - 2026
DA  - 2026/10/09
JO  - Adipocyte
DO  - 10.1080/21623945.2026.2736340
UR  - https://europepmc.org/article/MED/42852735
AB  - In vitro modelling of adipose tissue (AT) commonly employs 2D systems, which do not fully capture the complexity of AT, lacking cell-cell interactions and extracellular matrix (ECM) remodelling. Despite the innovations in 3D culture systems offering physiologically relevant AT platforms, the field lacks consensus and cross-protocol method comparison. We performed a comparative analysis of three scaffold-free spheroid-generation techniques: Ultra Low Attachment plates (ULA), Hanging Drops (HD), and Microwell Culture Plates (MCP), utilizing human AT-derived mesenchymal stromal cells. Each method produced viable spheroids of similar size and morphology within 48 hours. However, MCP demonstrated greater uniformity and a significant increase in mass density when compared to ULA and HD. Adipogenic media increased spheroid size in ULA and MCP, but not in HD spheroids. Notably, the diameter of ULA spheroids was 2- and 6-times larger compared to HD and MCP ones, respectively, regardless of adipogenic induction. Gene expression analysis indicated that all techniques led to PPARγ and ADIPOQ expression during adipogenesis. All methods supported adipocyte differentiation, as evidenced by lipid droplet accumulation, PLIN1, CD36, and HSL expression, and ECM rearrangement. These findings suggest that all the described techniques constitute a robust platform for producing adipogenesis-competent spheroids, while method-specific differences suggest an application-dependent choice.
SN  - 2162-3945
SN  - 2162-397X
ER  -