A comparative study of the capacity of mesenchymal stromal cell lines to form spheroids

Abstract

Mesenchymal stem cells (MSC)-spheroid models favor maintenance of stemness, ex vivo expansion and transplantation efficacy. Spheroids may also be considered as useful surrogate models of the hematopoietic niche. However, accessibility to primary cells, from bone marrow (BM) or adipose tissues, may limit their experimental use and the lack of consistency in methods to form spheroids may affect data interpretation. In this study, we aimed to create a simple model by examining the ability of cell lines, from human (HS-27a and HS-5) and murine (MS-5) BM origins, to form spheroids, compared to primary human MSCs (hMSCs). Our protocol efficiently allowed the spheroid formation from all cell types within 24 hours. Whilst hMSC-spheroids began to shrink after 24 hours, the size of spheroids from cell lines remained constant during three weeks. The difference was partially explained by the balance between proliferation and cell death, which could be triggered by hypoxia and induced oxidative stress. Our results demonstrate that, like hMSCs, MSC cell lines make reproductible spheroids that are easily handled. Thus, this model could help in understanding mechanisms involved in MSC functions and may provide a simple model by which to study cell interactions in the BM niche.

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

“…To further investigate the differences in cell number and to obtain spatial information about the location of dead cells, we performed a live/dead assay (Figure 3). We observed an increase in cell death on day 7 compared to day 1 in all culture systems, which was consistent with our DNA content result as well as other studies that have shown increase in cell death due to apoptosis when MSCs are cultured as aggregates under static conditions (Deynoux et al, 2020).…”

Mkx Mediates Tenogenic Differentiation But Incompletely Inhibits The Proliferation of Hypoxic MSCs

“…Many studies have reported that hypoxia prompted the self-renewal of MSCs [8][9][10][11]. This was consistent with our ndings which showed that the signi cantly higher OD values and clone numbers were detected in both AMSCs and BMSCs in the hypoxia group, compared with those in the normoxia group.…”

“…However, differentiation-inducing therapy of MSCs has been reported to be potential in treating tendon injury [2,6,7]. Among the various differentiation-inducing therapies, hypoxia has not only been found to promote the proliferation of MSCs [8][9][10][11], but also has shown to be an e cient inductor in tenogenic differentiation of MSCs in repairing tendon injury [8,12,13]. Many studies have found that hypoxia increased the number of MSCs and augmented the formation of colonies [8,9].…”

Impact of Bioreactor Geometry on Mesenchymal Stem Cell Production in Stirred‐Tank Bioreactors

“…The spheroids are continuously rearranged and condensed 18. This is supported by experimental data that we and others have published 27 showing that spheroid size declines with cultivation time. Others have reported hMSC spheroid growth with cultivation time 28, but this also leads to an increasing number of single cells or smaller cell agglomerates in the supernatant, which can form new spheroids.…”