A comparative study of mesenchymal stem cells cultured as cell‐only aggregates and in encapsulated hydrogels

Abstract

There is increasing evidence that cells cultured in three-dimensional (3D) settings have superior performance compared to their traditional counterparts in monolayers. This has been attributed to cell-cell and cell-extracellular matrix interactions that more closely resemble the in vivo tissue architecture. The rapid adoption of 3D cell culture systems as experimental tools for diverse applications has not always been matched by an improved understanding of cell behavior in different 3D environments. Here, we studied human mesenchymal stem/stromal cells (hMSCs) as scaffold-free self-assembled aggregates of low and high cell number and compared them to cell-laden alginate hydrogels with and without arginine-glycine-aspartic acid peptides. We observed a significant decrease in the size of cell-only aggregates over 14 days in culture compared to the cells encapsulated in alginate hydrogels. Alginate hydrogels had persistently more living cells for a longer period (14 days) in culture as measured by total DNA content. Proliferation studies revealed that a weeklong culture of hMSCs in 3D culture, whether as aggregates or cell-laden alginate hydrogels, reduced their proliferation over time.Cell cycle analysis found no significant differences between days 1 and 7 for the different culture systems. The findings of this study improve our understanding of how aggregate cultures differ with or without a hydrogel carrier, and whether aggregation itself is important when it comes to the 3D culture of hMSCs.

Alginate Core–Shell Capsules for 3D Cultivation of Adipose-Derived Mesenchymal Stem Cells

“…Most importantly, the capsule system did allow for cell survival and proliferation. An attenuated proliferation had been previously reported for MSCs maintained in spheroids [ 37 , 38 ], but next to maintaining a high viability, we also confirmed actively proliferating cells after the initial condensation phase. A 2.5-fold increase in cell numbers was achieved over the four days of cultivation.…”

“…Interestingly, in both groups, the signal shows a four–five-fold increase until days 2–3 but stays at that level or even decreases until day 4 ( Figure 6 C). There have been reports of MSCs cultured in spheroids attenuate proliferating after compacting [ 37 , 38 ]. Therefore, to investigate the proliferative behavior of the aggregated cells after this initial compaction phase, we performed EdU proliferation staining on days 2 and 4.…”

The Role of Microsphere Structures in Bottom-Up Bone Tissue Engineering

“…Additionally, the 3D culture of stem cells includes self-assembly aggregation and encapsulation by hydrogel microspheres. Passanha et al found that cells encapsulated by alginate hydrogel exhibited better viability on the 14th day, probably because the alginate gel could provide more room for cells and allow for the diffusion of more nutrients and oxygen [ 197 ].…”

Modular mixing of benzene-1,3,5-tricarboxamide supramolecular hydrogelators allows tunable biomimetic hydrogels for control of cell aggregation in 3D

“…After illustrating spheroid fusion in these hydrogels, we subsequently investigated hMSC proliferation capacity within the different formulations. Previous studies have shown that proliferation rates are different in 2D and 3D, and many cell lines show reduced proliferation in 3D spheroid cultures 62 and hydrogels (synthetic and natural) [65][66][67] compared to 2D monolayer cultures. The type of matrix 66 and stress relaxation 18 also have been shown to influence the proliferation rates for cells.…”