![]() ![]() Thus, biomaterial scaffolds that degrade slowly over months to years are required. Further, our long-term goal is a sustainable coculture system, such that the three-dimensional (3D) vascularized tissue can be utilized as sustainable disease models in vitro, or for long-term soft tissue repairs in vivo. 16 While these studies, among others, show promise toward vascularized engineered tissue, we are interested in establishing a functional model such that adipose-derived stromal cells are differentiated toward adipose tissue specifically, cocultured with endothelial cells on a material that has shown promise for the development of adipose tissue, and exhibit adipose tissue functionality. cocultured preadipocytes from human adipose tissue with endothelial cells in a fibrin glue matrix on the chorioallantoic membrane, and after 7 days culture on the CAM, positive CD31 stain was evident in lumen-like structures in the fibrin matrix. developed an in vitro coculture model using human adipose stromal cells and human umbilical vein endothelial cells (HUVECs), where perfused tubes demonstrated capillary-like networks that were observed sprouting from the central lumen wall. 4, 10– 13 Coculture models for adipose tissue engineering have been explored to a limited extent in prior reports. 9Ĭurrent in vitro adipose tissue models involve monocultures of differentiated bone marrow–derived mesenchymal stem cells, differentiated adipose tissue–derived stem cells, as well as embryonic stem cells cultured on synthetic or natural polymeric matrices or scaffolds. 8 Incorporating vasculature in vitro would contribute to viability during tissue growth and induce structural organization in vitro as well as in vivo, as well as provide more relevance to in vitro models of tissues. 7 Therefore, a current need in tissue engineering rests with the formation of a vascular supply in tissue-engineered constructs. However, a major obstacle to this approach is the lack of a vascular supply. One approach for an adipose tissue model is to tissue engineer adipose tissue in vitro to mimic native tissue. 6 These adipokines include leptin, adiponectin, tumor necrosis factor-α, plasminogen activator inhibitor type 1, and resistin. 4, 5 Recent studies have shown that a close correlation exists between adipogenesis and angiogenesis in which specific adipokines, or cytokines secreted by adipocytes, play a role in vascular homeostasis. 3 White adipose tissue, the predominant type of fat in humans, is composed of preadipocytes, differentiated adipocytes, interstitial cells, and a microvascular system that is organized within an extracellular matrix that consists of collagen types I, III, IV, V, and VI as well as other extracellular matrix proteins. ![]() 1 Obesity has severe consequences, such as increased risk of coronary heart disease, type 2 diabetes, and hypertension. ![]() According to the National Center for Health Statistics, 30% of adults in the United States are considered obese. These models can serve to elucidate mechanisms of disease origin and progression, as well as for therapeutic screening and diagnostic tools. The increased incidence of adipose-related diseases such as obesity and insulin resistance associated with type 2 diabetes prompts rising interest in the development of models to allow the more systematic study of disease mechanisms. The strategy outlined provides a basis for the formation of other in vitro vascularized tissues as well as a path forward for the sustainable formation of soft tissue due to the use of slowly degrading silk scaffolds.Ī n in vitro model of adipose tissue that mimics aspects of the native tissue would offer major benefits to the biomedical research community. A promising in vitro approach for the vascularization of tissue-engineered adipose tissue, and the ability to vascularize a construct containing hASCs was demonstrated. Additionally, lipid accumulation was demonstrated with Oil Red O staining, where positive staining was higher in the differentiated cocultures. Differentiated adipose cocultures secreted significantly higher levels of leptin than undifferentiated cocultures at 1 and 2 weeks. Endothelial cells aligned with time, and further histological analyses revealed continuous endothelial lumen formation in both differentiated and undifferentiated cocultures. Confocal microscopy images demonstrated viability of cocultures and organization of both cell types over time. Tomato red–infected human adipose tissue–derived mesenchymal stem cells (hASCs) and green fluorescence protein–infected human umbilical vein endothelial cells were cocultured on 3D aqueous-derived silk scaffolds for 2 weeks. In this study, we evaluated structural and functional characteristics of an in vitro 3D coculture model of vascularized adipose tissue. The clinical need for both three-dimensional (3D) soft tissue replacements and in vitro adipose tissue models continues to grow. ![]()
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