Abstract
BackgroundAdipose-derived mesenchymal stem cells (MSCs) have been gaining fame mainly due to their vast clinical potential, simple isolation methods and minimal donor site morbidity. Adipose-derived MSCs and microvascular endothelial cells have been shown to bear angiogenic and vasculogenic capabilities. We hypothesized that co-culture of human adipose-derived MSCs with human adipose-derived microvascular endothelial cells (HAMECs) will serve as an effective cell pair to induce angiogenesis and vessel-like network formation in three-dimensional scaffolds in vitro.MethodsHAMECs or human umbilical vein endothelial cells (HUVECs) were co-cultured on scaffolds with either MSCs or human neonatal dermal fibroblasts. Cells were immunofluorescently stained within the scaffolds at different time points post-seeding. Various analyses were performed to determine vessel length, complexity and degree of maturity.ResultsThe HAMEC:MSC combination yielded the most organized and complex vascular elements within scaffolds, and in the shortest period of time, when compared to the other tested cell combinations. These differences were manifested by higher network complexity, more tube alignment and higher α-smooth muscle actin expression. Moreover, these generated microvessels further matured and developed during the 14-day incubation period within the three-dimensional microenvironment.ConclusionsThese data demonstrate optimal vascular network formation upon co-culture of microvascular endothelial cells and adipose-derived MSCs in vitro and constitute a significant step in appreciation of the potential of microvascular endothelial cells and MSCs in different tissue engineering applications that can also be advantageous in in vivo studies.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0251-6) contains supplementary material, which is available to authorized users.
Highlights
Adipose-derived mesenchymal stem cells (MSCs) have been gaining fame mainly due to their vast clinical potential, simple isolation methods and minimal donor site morbidity
Vessel-like network development on poly-L-lactic acid (PLLA)/poly-lactic-coglycolic acid (PLGA) constructs In an effort to comprehend how vessel networks develop during in vitro culture, fluorescently stained Human adipose microvascular endothelial cell (HAMEC) and Human umbilical vein endothelial cell (HUVEC) grown in various co-culture combinations on 3D PLLA/PLGA matrices were tracked over a 14-day period using confocal microscopy
Well-developed vessellike networks of HAMECs:MSCs were already observed on day 7 (Fig. 1a), and were maintained throughout the remainder of the experiment
Summary
Adipose-derived mesenchymal stem cells (MSCs) have been gaining fame mainly due to their vast clinical potential, simple isolation methods and minimal donor site morbidity. Adipose-derived MSCs and microvascular endothelial cells have been shown to bear angiogenic and vasculogenic capabilities. We hypothesized that co-culture of human adipose-derived MSCs with human adipose-derived microvascular endothelial cells (HAMECs) will serve as an effective cell pair to induce angiogenesis and vessel-like network formation in three-dimensional scaffolds in vitro. Due to the capacity of its mesenchymal stem cell (MSC) content to differentiate to a variety of cell types of the mesodermal lineage [8, 9], recent tissue engineering applications have integrated adipose-derived MSCs to achieve regeneration of a variety of tissue types, including vascular tissues [10, 11]. The origin of ECs selected for vascular formation applications has been suggested to be very important as well, where adipose-derived microvascular ECs have been shown to display stronger angiogenic capacities, when compared to those derived from macrovessels [13, 14]
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