Department of Chemical Engineering, Virginia Tech, Blacksburg, Virginia 24061, United States.
ACS Biomater Sci Eng. 2023 Jun 12;9(6):3445-3461. doi: 10.1021/acsbiomaterials.2c01310. Epub 2023 May 2.
Glioblastoma multiforme (GBM) is the deadliest form of brain cancer, responsible for over 50% of adult brain tumors. A specific region within the GBM environment is known as the perivascular niche (PVN). This area is defined as within approximately 100 μm of vasculature and plays an important role in the interactions between endothelial cells (ECs), astrocytes, GBM cells, and stem cells. We have designed a 3D model of the PVN comprising either collagen Type 1 or HyStem-C, human umbilical vein ECs (HUVECs), and LN229 (GBM) cells. HUVECs were encapsulated within the hydrogels to form vascular networks. After 7 days, LN229 cells were co-cultured to investigate changes in both cell types. Over a 14 day culture period, we measured alterations in HUVEC networks, the contraction of the hydrogels, trans-differentiation of LN229 cells, and the concentrations of two chemokines; CXCL12 and TGF-β. Increased cellular proliferation ranging from 10- to 16-fold was exhibited in co-cultures from days 8 to 14. This was accompanied with a decrease in the height of hydrogels of up to 68%. These changes in the biomaterial scaffold indicate that LN229-HUVEC interactions promote changes to the matrix. TGF-β and CXCL12 secretion increased approximately 2-2.6-fold each from day 8 to 14 in all co-cultures. The expression of CXCL12 correlated with cell colocalization, indicating a chemotactic role in enabling the migration of LN229 cells toward HUVECs in co-cultures. von Willebrand factor (vWF) was co-expressed with glial fibrillary acidic protein (GFAP) in up to 15% of LN229 cells after 24 h in co-culture. Additionally, when LN229 cells were co-cultured with human brain microvascular ECs, the percentages of GFAP/vWF cells were up to 20% higher than that in co-cultures with HUVECs in collagen (2.2 mg/mL) and HyStem-C gels on day 14. The expression of vWF indicates the early stages of trans-differentiation of LN229 cells to an EC phenotype. Designing models of trans-differentiation may provide additional insights into how vasculature and cellular phenotypes are altered in GBM.
多形性胶质母细胞瘤（GBM）是最致命的脑癌形式，占成人脑肿瘤的 50%以上。GBM 环境中的一个特定区域称为血管周龛（PVN）。该区域定义为血管周围约 100μm 的范围，在血管内皮细胞（ECs）、星形胶质细胞、GBM 细胞和干细胞之间的相互作用中起着重要作用。我们设计了一种包含胶原 1 型或 HyStem-C、人脐静脉内皮细胞（HUVECs）和 LN229（GBM）细胞的 PVN 的 3D 模型。HUVECs 被包封在水凝胶中以形成血管网络。7 天后，共培养 LN229 细胞以研究两种细胞类型的变化。在 14 天的培养期间，我们测量了 HUVEC 网络的改变、水凝胶的收缩、LN229 细胞的转分化以及两种趋化因子 CXCL12 和 TGF-β的浓度。从第 8 天到第 14 天，共培养物中的细胞增殖增加了 10 到 16 倍。这伴随着水凝胶高度降低了高达 68%。生物材料支架的这些变化表明 LN229-HUVEC 相互作用促进了基质的变化。TGF-β和 CXCL12 的分泌从第 8 天到第 14 天在所有共培养物中增加了约 2-2.6 倍。CXCL12 的表达与细胞共定位相关，表明在共培养物中 LN229 细胞向 HUVEC 迁移的趋化作用。von Willebrand 因子（vWF）在共培养 24 小时后与神经胶质纤维酸性蛋白（GFAP）在高达 15%的 LN229 细胞中共表达。此外，当 LN229 细胞与人脑微血管内皮细胞共培养时，GFAP/vWF 细胞的百分比在第 14 天胶原（2.2mg/ml）和 HyStem-C 凝胶中的共培养物中比 HUVEC 高 20%。vWF 的表达表明 LN229 细胞向 EC 表型的早期转分化。设计转分化模型可能为研究血管生成和细胞表型在 GBM 中的改变提供更多的见解。
