Long term morphological characterization of mesenchymal stromal cells 3D spheroids built with a rapid method based on entry-level equipment

Bellotti, Chiara; Duchi, Serena; Bevilacqua, Alessandro; Lucarelli, Enrico; Piccinini, Filippo

doi:10.1007/s10616-016-9969-y

Cited by 31 publications

(24 citation statements)

References 48 publications

(57 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, HS-27a showed a decrease in viable cells similarly to primary hMSCs and MS-5 are quiescent but do not shrink. Shrinking has already been reported for primary hMSCs [31,32,35,37,[67][68][69] and has been attributed to autophagy [32]. Reduced cell growth or arrest is known to trigger autophagy [70].…”

Section: Plos Onementioning

confidence: 90%

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

et al. 2020

View full text Add to dashboard Cite

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.

show abstract

Section: Plos Onementioning

confidence: 90%

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

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Spheroid areas were determined using ImageJ software Version 1.6. Starting from the binary masks obtained by Image J, the volume of each spheroid was computed using ReViSP [50], a software specifically designed to accurately estimate the volume of spheroids and to render an image of their 3D surface.…”

Section: Methodsmentioning

confidence: 99%

Comparison between adult and foetal adnexa derived equine post-natal mesenchymal stem cells

et al. 2019

View full text Add to dashboard Cite

Background Little is known about the differences among adult and foetal equine mesenchymal stem cells (MSCs), and no data exist about their comparative ultrastructural morphology. The aim of this study was to describe and compare characteristics, immune properties, and ultrastructural morphology of equine adult (bone marrow: BM, and adipose tissue: AT) and foetal adnexa derived (umbilical cord blood: UCB, and Wharton’s jelly: WJ) MSCs. Results No differences were observed in proliferation during the first 3 passages. While migration ability was similar among cells, foetal MSCs showed a higher adhesion ability, forming smaller spheroids after hanging drop culture ( P < 0.05). All MSCs differentiated toward adipogenic, chondrogenic and osteogenic lineages, only tenogenic differentiation was less evident for WJ-MSCs. Data obtained by PCR confirmed MHC1 expression and lack of MHC2 expression in all four cell types. Foetal adnexa MSCs were positive for genes specific for anti-inflammatory and angiogenic factors (IL6, IL8, ILβ1) and WJ-MSCs were the only positive for OCT4 pluripotency gene. At immunofluorescence all cells expressed typical mesenchymal markers (α-SMA, N-cadherin), except for BM-MSCs, which did not express N-cadherin. By transmission electron microscopy, it was observed that WJ-MSCs had a higher ( P < 0.05) number of microvesicles compared to adult MSCs, and UCB-MSCs showed more microvesicles than BM-MSCs ( P < 0.05). AT-MSCs had a lower number of mitochondria than WJ-MSCs ( P < 0.05), and mitochondrial area was higher for WJ-MSCs compared to UCB and AT-MSCs ( P < 0.05). Conclusions Results demonstrate that MSCs from adult and foetal tissues have different characteristics, and foetal MSCs, particularly WJ derived ones, seem to have some charactestics that warrant further investigation into potential advantages for clinical application.

show abstract

“…Spheroid areas were determined using ImageJ software (imagej.nih.gov/ij/). Starting from the binary masks obtained by ImageJ, the volume of each spheroid was computed using ReViSP (sourceforge.net/ projects/revisp) (Bellotti et al 2016), a software specifically designed to accurately estimate the volume of spheroids and to render an image of their 3D surface.…”

Section: Adhesion and Migration Assaysmentioning

confidence: 99%

Ultrastructural characteristics and immune profile of equine MSCs from fetal adnexa

Iacono¹,

Pascucci²,

Rossi³

et al. 2017

Reproduction

View full text Add to dashboard Cite

Both in human and equine species, mesenchymal stem cells (MSCs) from amniotic membrane (AM) and Wharton's jelly (WJ), may be particularly useful for immediate use or in later stages of life, after cryopreservation in cell bank. The aim of this study was to compare equine AM- and WJ-MSCs features that may be relevant for their clinical employment. MSCs were more easily isolated from WJ, even if MSCs derived from AM exhibited more rapid proliferation ( < 0.05). Osteogenic and chondrogenic differentiation were more prominent in MSCs derived from WJ. This is also suggested by the lower adhesion of AM cells, demonstrated by the greater volume of spheroids after hanging drop culture ( < 0.05). Data obtained by PCR confirmed the immunosuppressive function of AM and WJ-MSCs and the presence of active genes specific for anti-inflammatory and angiogenic factors (IL-6, IL 8, IL-β1). For the first time, by means of transmission electron microscopy (TEM), we ascertained that equine WJ-MSCs constitutively contain a very impressive number of large vesicular structures, scattered throughout the cytoplasm. Moreover, an abundant extracellular fibrillar matrix was located in the intercellular spaces among WJ-MSCs. Data recorded in this study reveal that MSCs from different fetal tissues have different characteristics that may drive their therapeutic use. These finding could be noteworthy for horses as well as for other mammalian species, including humans.

show abstract

Long term morphological characterization of mesenchymal stromal cells 3D spheroids built with a rapid method based on entry-level equipment

Cited by 31 publications

References 48 publications

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

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

Comparison between adult and foetal adnexa derived equine post-natal mesenchymal stem cells

Ultrastructural characteristics and immune profile of equine MSCs from fetal adnexa

Contact Info

Product

Resources

About