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Lipolytic function of adipocyte/endothelial cocultures.

ABSTRACT: The rising incidence of adipose-related disorders such as obesity has prompted increased interest in the in vitro development of functional human soft tissues to study the disease and treatment options. Further, soft tissues maintained in vitro with a capacity to resemble in vivo tissues in structure and metabolic function would help gain insight into mechanisms involved in adipose tissue development. In the current study, the metabolic potential of adipose/endothelial cocultures on three-dimensional silk fibroin scaffolds was studied. Endothelial contributions to adipose lipogenesis and lipolysis were the focus of the study. Triglyceride accumulation, adipogenic gene transcript expression, and basal lipolysis measurements demonstrated the ability of this coculture system to retain metabolic levels obtained in adipocyte monocultures. Additionally, basal lipolysis was stimulated in mono- and coculture systems to a similar extent at 1.6- and 1.9-fold over controls, respectively. The ability to maintain adipose functions in these cocultures represents a step forward in the development of a tissue-engineered adipose tissue system exhibiting both endothelial lumens and metabolic functions.

SUBMITTER: Choi JH

PROVIDER: S-EPMC3079167 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature

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Lipolytic function of adipocyte/endothelial cocultures.

Choi Jennifer H JH Bellas Evangelia E Gimble Jeffrey M JM Vunjak-Novakovic Gordana G Kaplan David L DL

Tissue engineering. Part A 20110303 9-10

The rising incidence of adipose-related disorders such as obesity has prompted increased interest in the in vitro development of functional human soft tissues to study the disease and treatment options. Further, soft tissues maintained in vitro with a capacity to resemble in vivo tissues in structure and metabolic function would help gain insight into mechanisms involved in adipose tissue development. In the current study, the metabolic potential of adipose/endothelial cocultures on three-dimens ...[more]

PMID: 21247356

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Project description:The development of viable tissue surrogates requires a vascular network that sustains cell metabolism and tissue development. The coculture of endothelial cells (ECs) and mesenchymal stem cells (MSCs), the two key players involved in blood vessel formation, has been heralded in tissue engineering (TE) as one of the most promising approaches for scaffold vascularization. However, MSCs may exert both proangiogenic and antiangiogenic role. Furthermore, it is unclear which cell type is responsible for the upregulation of angiogenic pathways observed in EC:MSC cocultures. There is disagreement on the proangiogenic action of MSCs, as they have also been shown to negatively affect the formation of capillary networks. To address these issues, we investigated the regulation of key angiogenic pathways in scaffolds hosting different EC:MSC ratios fabricated through extrusion-based bioprinting. Human ECs were cocultured with either rat or human MSCs, and the regulation of fundamental angiogenic and arteriogenic pathways was analyzed through DNA, gene, and protein expression. The use of a hybrid human/rat coculture system facilitated pinpointing each cell type role in the regulation of specific genes and showed that MSCs exert a dose-dependent inhibitory effect on the EC expression of angiogenic factors within the first 24 h. Within a week of coculture, MSCs exert a proangiogenic effect, as corroborated in human/human bioprinted cocultures. Interestingly, juxtacrine signaling promoted secretion of the angiogenic factor vascular endothelial growth factor in direct cocultures (EC and MSC co-encapsulated), while paracrine signaling encouraged secretion of the arteriogenic factor platelet-derived growth factor in indirect cocultures (adjacent bioprinting of EC-laden and MSC-laden scaffolds). Overall, the use of a bioprinted system to elucidate EC:MSC interplay allows rapid leveraging of the data for novel vascular TE applications. Despite the transitory negative effect early in the culture, MSC presence is necessary for the regulation of pathways involved in arteriogenesis. With further validation in vivo, this study provides a possible explanation to the controversial findings present in literature and shows how MSC effect on angiogenic pathway regulation mimics the dynamics of blood vessel formation reported in literature and normally occurring in vivo. Impact Statement The coculturing of endothelial cells (ECs) and mesenchymal stem cells (MSCs) holds great promise in tissue engineering for the development of prevascularized tissue constructs. Yet, different studies report conflicting results on the role of MSCs, which can either support or inhibit vasculature formation. Furthermore, it is unclear how each cell type modulates distinct pathways involved in angiogenesis when cocultured. Using bioprinted hybrid coculture systems, we show that MSCs have both a time- and dose-dependent effect on the gene and protein expression of key angiogenic and arteriogenic factors by ECs. These findings, obtained in translationally relevant setup, can readily inform the design of vascularized scaffolds.

Dataset Information

Lipolytic function of adipocyte/endothelial cocultures.

Publications

Lipolytic function of adipocyte/endothelial cocultures.

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