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Spheroids from adipose‐derived stem cells exhibit an miRNA profile of highly undifferentiated cells
Authors:A Barbara Di Stefano PhD  Federica Grisafi MSc  Marta Castiglia PhD  Alessandro Perez PhD  Luigi Montesano MD  Alessandro Gulino PhD  Francesca Toia MD  Daniele Fanale PhD  Antonio Russo MD  Francesco Moschella MD  Angelo A Leto Barone MD  Adriana Cordova MD
Affiliation:1. Department of Surgical, Oncological and Oral Sciences, Section of Plastic and Reconstructive Surgery, University of Palermo, Palermo, Italy;2. Department of Surgical, Oncological and Oral Sciences, Section of Medical Oncology, University of Palermo, Palermo, Italy;3. Department of Health Science, Human Pathology Section, Tumor Immunology Unit, University of Palermo, Palermo, Italy;4. Department of Plastic and Reconstructive Surgery, Johns Hopkins School of Medicine, Baltimore, MD
Abstract:Two‐dimensional (2D) cell cultures have been extensively used to investigate stem cell biology, but new insights show that the 2D model may not properly represent the potential of the tissue of origin. Conversely, three‐dimensional cultures exhibit protein expression patterns and intercellular junctions that are more representative of their in vivo condition. Multiclonal cells that grow in suspension are defined as “spheroids,” and we have previously demonstrated that spheroids from adipose‐derived stem cells (S‐ASCs) displayed enhanced regenerative capability. With the current study, we further characterized S‐ASCs to further understand the molecular mechanisms underlying their stemness properties. Recent studies have shown that microRNAs (miRNAs) are involved in many cellular mechanisms, including stemness maintenance and proliferation, and adipose stem cell differentiation. Most studies have been conducted to identify a specific miRNA profile on adherent adipose stem cells, although little is still known about S‐ASCs. In this study, we investigate for the first time the miRNA expression pattern in S‐ASCs compared to that of ASCs, demonstrating that cell lines cultured in suspension show a typical miRNA expression profile that is closer to the one reported in induced pluripotent stem cells. Moreover, we have analyzed miRNAs that are specifically involved in two distinct moments of each differentiation, namely early and late stages of osteogenic, adipogenic, and chondrogenic lineages during long‐term in vitro culture. The data reported in the current study suggest that S‐ASCs have superior stemness features than the ASCs and they represent the true upstream stem cell fraction present in adipose tissue, relegating their adherent counterparts.
Keywords:adipose stem cells  long‐term culture  mesenchymal differentiation  miR‐142‐3p  miRNAs
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