Preparation and characterization of directed, one‐day‐self‐assembled millimeter‐size spheroids of adipose‐derived mesenchymal stem cells

Abstract Three‐dimensional cell spheroids prepared without using any artificial scaffold materials are desirable for cell‐based transplants. However, conventional cell culture systems are inefficient for rapid, large‐scale and non‐cytotoxic generation of size‐controlled spheroids (>1 mm diameter) that are required for tissue regenerative therapy application. In this study, we prepared millimeter‐order spheroids of adipose‐derived mesenchymal stem cells (ADSCs) by controlling the spheroid size (diameter range: 0.4–2.5 mm). Notably, spheroid generation required only one day of culture on charged culture dishes. Almost all spheroid‐derived ADSCs were viable and produced adhesion molecules and growth factors, which play an important role in tissue regeneration. Moreover, spheroid‐derived ADSCs could infiltrate and recellularize collagenous tissue membranes in vitro . The ADSC spheroids developed in this study could be directly (without additional processing) used for cell‐based tissue regeneration therapy. Furthermore, the rapid scale‐up process and noncytotoxic generation of spheroids would also enable other applications such as use as screening models for drug discovery. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 305–312, 2016.