Generation of Three-Dimensional Fat-Glioblastoma Assembloid Systems
Büşra Yüksel, Mine Pelin Tansi, Ayşegül DoğanPURPOSE
Glioblastoma (GBM) is a highly heterogeneous brain tumor with limited treatment options and poor prognosis․ Current in vitro research is typically done in artificially condensed two-dimensional culture systems‚ which do not recapitulate the tumor microenvironment․ Thus‚ this study intended to create a new 3D fat-glioblastoma assembloid model to mimic tumor-microenvironment interactions more accurately while providing a more physiologically relevant platform to study cancer biology․
METHODS
This study aims to establish three-dimensional fat–glioblastoma assembloid systems by co-culturing glioblastoma spheroids with white adipose tissue (WAT) organoids. Glioblastoma spheroids will be generated from U87MG and IDH-mutant U87 cell lines using 3D culture systems. WAT organoids will be derived from human induced pluripotent stem cells (iPSCs) through stepwise differentiation into neuromesodermal progenitors (NMPs) and mesodermal lineages, followed by adipogenic induction. Cells will be cultured within an ECM-based system to promote adipose tissue maturation. WAT organoids will be characterized using Oil Red O staining and hematoxylin staining to confirm lipid accumulation and cellular morphology, along with gene expression analysis (qRT-PCR) and immunofluorescence. Following co-culture, assembloid systems will be established to investigate tumor–adipose interactions. Assembloid characterization will be performed using qRT-PCR and immunofluorescence, together with confocal microscopy to assess protein expression, cellular organization, and microenvironmental interactions.
RESULTS
Despite being under development‚ initial results suggest that glioblastoma spheroids and adipose-derived structures can be assembled into an asembloid․ It is not yet fully characterized for morphology and molecular properties‚ but it is already usable for inspecting how the tumor interacts with the microenvironment․
CONCLUSION
This study aims toward the development of a 3D asembloid model that better recapitulates the glioblastoma microenvironment and gives a better understanding of tumor-microenvironment interactions with the goal of providing more clinically predictive preclinical models․ In the long term‚ this study will ease more personalized therapy approaches․