Use of Culture Geometry to Control Hypoxia-Induced Vascular Endothelial Growth Factor Secretion from Adipose-Derived Stem Cells: Optimizing a Cell-Based Approach to Drive Vascular Growth

Skiles, Matthew L.; Sahai, Suchit; Rucker, Lindsay; Blanchette, James O.

doi:10.1089/ten.tea.2012.0750

Cited by 45 publications

(36 citation statements)

References 42 publications

(41 reference statements)

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“…In excessively large constructs, limited diffusion of oxygen, nutrients and metabolites may compromise cell survival, resulting in the formation of necrotic cores 5 . On the other hand, spheroids larger than 100 µm were reported to exhibit increased VEGF secretion, as compared to the smaller ones, so the selection of spheroids size should be a well-thought-out process 38 . MSC-OEC spheroids, generated both under control and XF conditions, were characterized by a decrease in metabolic activity at the onset of culture.…”

Section: Discussionmentioning

confidence: 99%

Xeno-free pre-vascularized spheroids for therapeutic applications

Bauman

Feijão

Carvalho

et al. 2018

Sci Rep

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Spheroid culture has gained increasing popularity, arising as a promising tool for regenerative medicine applications. Importantly, spheroids may present advantages over single-cell suspensions in cell-based therapies (CT). Unfortunately, most growth media used for spheroid culture contain animal origin-components, such as fetal bovine serum (FBS). The presence of FBS compromises the safety of CT and presents economic and ethical constraints. SCC (supplement for cell culture) is a novel xeno-free (XF) industrial cell culture supplement, derived from well-controlled pooled human plasma and processed under good manufacturing practice rules. Here, we developed a XF SCC-based formulation for 2D-culture of outgrowth endothelial cells (OEC), and then used it for generating co-culture spheroids of OEC and mesenchymal stem cells (MSC). XF MSC-OEC spheroids were characterized in detail and compared to spheroids cultured in FBS-supplemented medium. XF spheroids presented comparable integrity, size and morphology as the reference culture. The use of both media resulted in spheroids with similar structure, abundant extracellular matrix deposition and specific patterns of OEC distribution and organization. Notably, XF spheroids presented significantly enhanced angiogenic potential, both in vitro (fibrin sprouting assay) and in vivo (CAM assay). These findings are particularly promising in the context of potential therapeutic applications.

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Section: Discussionmentioning

confidence: 99%

Xeno-free pre-vascularized spheroids for therapeutic applications

Bauman

Feijão

Carvalho

et al. 2018

Sci Rep

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“…We evaluated the in vitro proangiogenic characteristics of thymus MSCs combined with HUVECs using a conventional 2D monolayer method and a quantitative 3D spheroid sprouting assay because 3D culture of MSCs has been demonstrated to potentiate their proangiogenic effects and may be more representative of their in vivo behavior [14,15].…”

Section: Proangiogenic Characteristics Of Thymus Mscsmentioning

confidence: 99%

Characterization and Angiogenic Potential of Human Neonatal and Infant Thymus Mesenchymal Stromal Cells

Wang

Mundada

Johnson

et al. 2015

Stem Cells Translational Medicine

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Mesenchymal stromal cells (MSCs) are present in large numbers in discarded neonatal and infant thymus tissue and can be isolated by an explant culture method. Explant culture‐isolated thymus MSCs demonstrate the classical characteristics of bone marrow MSCs. Thymus MSCs also possess proangiogenic qualities and deserve further consideration as a potential cell therapeutic agent to promote tissue regeneration and repair and for use in strategies to vascularize engineered tissues.

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“…Differences in secretion levels from ASC spheroids due to spheroid size have not been widely investigated, though size-dependent differences in degree and homogeneity of chondrogenic differentiation of MSC spheroids have been reported (29). Necrosis of cells within the center of large spheroids has also been reported, suggesting an explanation for the superior per-cell production from 10k spheroids (43,55). Regional differences in cell behavior, including potential phenotypic commitment and corresponding changes in gene expression, could help explain the lack of detectable production of other angiogenic factors that have been commonly reported, such as EGF, PDGF, FGF-2, and HGF.…”

Section: Discussionmentioning

confidence: 94%

ASC Spheroid Geometry and Culture Oxygenation Differentially Impact Induction of Preangiogenic Behaviors in Endothelial Cells

et al. 2015

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Cell-based angiogenic therapies offer potential for the repair of ischemic injuries, while avoiding several of the limitations associated with material-based growth factor delivery strategies. Evidence supports that applying MSCs as spheroids rather than dispersed cells can improve retention and enhance therapeutic effect through increased secretion of angiogenic factors due to hypoxia. However, while spheroid culture appears to modulate MSC behavior, there has been little investigation of how major culture parameters that affect cellular oxygen tension, such as external oxygenation and culture size, impact the angiogenic potential of spheroids. We cultured equal numbers of adipose-derived stem cells (ASCs) as spheroids containing 10,000 (10k) or 60,000 (60k) cells each, in 20% and 2% oxygen. VEGF secretion varied among the sample groups, with 10k, 2% O2 spheroids exhibiting the highest production. Spheroid-conditioned media was applied to HUVEC monolayers, and proliferation was assessed. Spheroids of either size in 2% oxygen induced comparable proliferation compared to a 2 ng/ml VEGF control sample, while spheroids in 20% oxygen induced less proliferation. Spheroids were also applied in coculture with HUVEC monolayers, and induction of migration through a Transwell membrane was evaluated. Sixty thousand, 2% O2 spheroids induced similar levels of migration as VEGF controls, while 10k, 2% O2 spheroids induced significantly more. Ten thousand, 20% spheroids performed no better than VEGF-free controls. We conclude that the therapeutic ability of ASC spheroids to stimulate angiogenesis in endothelial cells is affected by both culture size and oxygenation parameters, suggesting that, while ASC spheroids offer potential in the treatment of injured and ischemic tissues, careful consideration of culture size in respect to in vivo local oxygen tension will be necessary for optimal results.

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Use of Culture Geometry to Control Hypoxia-Induced Vascular Endothelial Growth Factor Secretion from Adipose-Derived Stem Cells: Optimizing a Cell-Based Approach to Drive Vascular Growth

Cited by 45 publications

References 42 publications

Xeno-free pre-vascularized spheroids for therapeutic applications

Xeno-free pre-vascularized spheroids for therapeutic applications

Characterization and Angiogenic Potential of Human Neonatal and Infant Thymus Mesenchymal Stromal Cells

ASC Spheroid Geometry and Culture Oxygenation Differentially Impact Induction of Preangiogenic Behaviors in Endothelial Cells

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