Details

Autor(en) / Beteiligte

• Amemiya, Takeo
• Hata, Nobuhiro
• Mizoguchi, Masahiro
• Yokokawa, Ryuji
• Kawamura, Yoichiro
• Hatae, Ryusuke
• Sangatsuda, Yuhei
• Kuga, Daisuke
• Fujioka, Yutaka
• Takigawa, Kosuke
• Akagi, Yojiro
• Yoshimoto, Koji
• Iihara, Koji
• Miura, Takashi

Titel

Mesenchymal glioblastoma-induced mature de-novo vessel formation of vascular endothelial cells in a microfluidic device

Ist Teil von

• Molecular biology reports, 2021-01, Vol.48 (1), p.395-403

Ort / Verlag

Dordrecht: Springer Netherlands

Erscheinungsjahr

2021

Quelle

MEDLINE

Beschreibungen/Notizen

• High vascularization is a biological characteristic of glioblastoma (GBM); however, an in-vitro experimental model to verify the mechanism and physiological role of vasculogenesis in GBM is not well-established. Recently, we established a self-organizing vasculogenic model using human umbilical vein endothelial cells (HUVECs) co-cultivated with human lung fibroblasts (hLFs). Here, we exploited this system to establish a realistic model of vasculogenesis in GBM. We developed two polydimethylsiloxane (PDMS) devices, a doughnut-hole dish and a 5-lane microfluidic device to observe the contact-independent effects of glioblastoma cells on HUVECs. We tested five patient-derived and five widely used GBM cell lines. Confocal fluorescence microscopy was used to observe the morphological changes in Red Fluorescent Protein (RFP)-HUVECs and fluorescein isothiocyanate (FITC)-dextran perfusion. The genetic and expression properties of GBM cell lines were analyzed. The doughnut-hole dish assay revealed KNS1451 as the only cells to induce HUVEC transformation to vessel-like structures, similar to hLFs. The 5-lane device assay demonstrated that KNS1451 promoted the formation of a vascular network that was fully perfused, revealing the functioning luminal construction. Microarray analysis revealed that KNS1451 is a mesenchymal subtype of GBM. Using a patient-derived mesenchymal GBM cell line, mature de-novo vessel formation could be induced in HUVECs by contact-independent co-culture with GBM in a microfluidic device. These results support the development of a novel in vitro research model and provide novel insights in the neovasculogenic mechanism of GBM and may potentially facilitate the future detection of unknown molecular targets.