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.

Monitoring ZEO apoptotic potential in 2D and 3D cell cultures and associated spectroscopic evidence on mode of interaction with DNA

“…Furthermore spheroid’s diameter is an important parameter in characterizing drug efficiency, for instance, small spheroids with diameters up to 200 μm are sufficient to reflect in vivo -like, cell-cell and cell-matrix interactions and are frequently used for anticancer drug screening 17 . However, spheroids with diameters of approximately 500 μm have been suggested to be a better choice than small spheroids, because they mimic the pathophysiological conditions of solid tumors, such as the specific hypoxic areas in the center and proliferation gradients more properly 18, 19 . For this reason, homogenous multicellular spheroids from MDA-MB-231 cells were generated by using Hanging Drop and subsequent maintenance and increase of spheroids size was achieved by using Liquid Overlay method 20 .…”

Osteochondral Regeneration Using Adipose Tissue-Derived Mesenchymal Stem Cells

“…The use of spheroid formation for cartilage and osteochondral regeneration using AT-MSCs has been reviewed in this section and summarized in Table 2. First, we introduced the preparation and characterization of directed, one-day-self-assembled millimeter-sized spheroids of AT-MSCs [57]. Second, enhanced cartilage formation via 3D cell engineering of AT-MSCs was introduced [58].…”

“…In this review, recent studies, demonstrating cartilage and osteochondral regeneration following implantation of AT-MSCs, were first introduced [48][49][50][51][52][53][54][55][56]. Second, the most recent examples of assembling AT-MSCs in three dimensions with or without scaffold for cartilage regeneration were discussed [57][58][59][60]. Third, osteochondral regeneration through implantation of scaffold-free constructs, consisting of autologous swine AT-MSCs, fabricated using the "mold", into osteochondral defects in minipigs was discussed [39,40].…”

Spheroids from adipose‐derived stem cells exhibit an miRNA profile of highly undifferentiated cells

“…Even though cells lose their structural characteristics when they are separated from the original tissue, there are different techniques that enable one to reestablish three-dimensionality and cell-cell contacts. A 3D environment stimulates cells to grow in suspended aggregates that are defined as "spheroids" (Di Stefano et al, 2015), and there are different available isolation methods, such as the spinner (Mueller-Klieser, 1987), hanging-drop (Naderi et al, 2014), and coated-supports (Iwai, Nemoto, & Nakayama, 2016, Suenaga, Furukawa, Suzuki, Takato, & Ushida, 2015 techniques.…”