The development of vascularized and functional adipose tissue substitutes is required to improve soft tissue augmentation. In this study, vascularized adipose tissue constructs were generated using uncultured cells from the stromal-vascular fraction (SVF) of adipose tissue as an alternative cell source to adipose-derived stem cells. SVF cell behavior and tissue formation were compared in a stable fibrin formulation developed by our group and a commercial fibrin sealant (TissuCol; Baxter) upon direct subcutaneous implantation in a nude mouse model. Further, the effect of in vitro adipogenic induction on SVF cell development was investigated by implanting stable fibrin constructs after 1 week of precultivation (adipogenic vs. noninduced control). Constructs were thoroughly analyzed before implantation regarding adipogenic differentiation status, cell viability, and distribution as well as the presence of endothelial cells. Before implantation, in vitro precultivation strongly promoted adipogenesis (under adipogenic conditions) and the formation of CD31(+) prevascular structures by SVF cells (under nonadipogenic conditions). Tissue development in vivo was determined after 4 weeks by histology (hematoxylin and eosin, human vimentin) and quantified histomorphometrically. In stable fibrin gels, adipogenic precultivation was superior to noninduced conditions, resulting in mature adipocytes and the formation of distinct vascular structures of human origin in vivo. Strong neovascularization by the implanted cells predominated in noninduced constructs. Without pretreatment, the SVF in stable fibrin gels displayed only a weak differentiation capability. In contrast, TissuCol gels strongly supported the formation of coherent and well-vascularized adipose tissue of human origin, displaying large unilocular adipocytes. The developed native-like tissue architecture was highlighted by a whole mount staining technique. Taken together, SVF cells from human adipose tissue were shown to successfully lead to adipose tissue formation in fibrin hydrogels in vivo. The results render the SVF a promising cell source for subsequent studies both in vitro and in vivo with the aim of engineering clinically applicable soft tissue substitutes.
BackgroundMedulloblastoma is the most common malignant brain tumor in children and can be divided in different molecular subgroups. Patients whose tumor is classified as a Group 3 tumor have a dismal prognosis. However only very few tumor models are available for this subgroup.MethodsWe established a robust orthotopic xenograft model with a cell line derived from the malignant pleural effusions of a child suffering from a Group 3 medulloblastoma.ResultsBesides classical characteristics of this tumor subgroup, the cells display cancer stem cell characteristics including neurosphere formation, multilineage differentiation, CD133/CD15 expression, high ALDH-activity and high tumorigenicity in immunocompromised mice with xenografts exactly recapitulating the original tumor architecture.ConclusionsThis model using unmanipulated, human medulloblastoma cells will enable translational research, specifically focused on Group 3 medulloblastoma.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2170-z) contains supplementary material, which is available to authorized users.
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