Department of Orthopedics, Technical University Munich, Munich, Germany.
Department of Urology, Technical University Munich, Munich, Germany.
Sci Rep. 2017 Jul 10;7(1):4991. doi: 10.1038/s41598-017-04572-1.
The growing interest in engineered tumor models prompted us to devise a method for the non-invasive assessment of such models. Here, we report on bioluminescence imaging (BLI) for the assessment of engineered tumor models in the fertilized chicken egg, i.e, chick chorioallantoic membrane (CAM) assay. One prostate cancer (PC-3) and two osteosarcoma (MG63 and HOS) cell lines were modified with luciferase reporter genes. To create engineered tumors, these cell lines were seeded either onto basement membrane extract (BME) or gelfoam scaffolds, and subsequently grafted in vivo onto the CAM. BLI enabled non-invasive, specific detection of the engineered tumors on the CAM in the living chicken embryo. Further, BLI permitted daily, quantitative monitoring of the engineered tumors over the course of up to 7 days. Data showed that an extracellular matrix (ECM) composed of BME supported growth of reporter gene marked PC-3 tumors but did not support MG63 or HOS tumor growth. However, MG63 tumors engineered on the collagen-based gelfoam ECM showed a temporal proliferation burst in MG63 tumors. Together, the data demonstrated imaging of engineered human cancer models in living chicken embryos. The combination of CAM assay and BLI holds significant potential for the examination of a broad range of engineered tumor models.
人们对工程化肿瘤模型越来越感兴趣,这促使我们设计了一种非侵入性评估此类模型的方法。在这里,我们报告了用于评估鸡胚中工程化肿瘤模型的生物发光成像 (BLI),即鸡胚绒毛尿囊膜 (CAM) 分析。我们对前列腺癌细胞系 (PC-3) 和两个成骨肉瘤细胞系 (MG63 和 HOS) 进行了荧光素酶报告基因修饰。为了构建工程化肿瘤,将这些细胞系接种到基底膜提取物 (BME) 或明胶海绵支架上,然后在体内移植到 CAM 上。BLI 能够在活体鸡胚中对 CAM 上的工程化肿瘤进行非侵入性、特异性检测。此外,BLI 允许在长达 7 天的时间内每天对工程化肿瘤进行定量监测。数据表明,由 BME 组成的细胞外基质 (ECM) 支持标记有报告基因的 PC-3 肿瘤的生长,但不支持 MG63 或 HOS 肿瘤的生长。然而,在基于胶原蛋白的明胶海绵 ECM 上构建的 MG63 肿瘤显示出 MG63 肿瘤的短暂增殖爆发。总之,这些数据证明了在活体鸡胚胎中对工程化人类癌症模型进行成像。CAM 分析和 BLI 的结合为广泛的工程化肿瘤模型的检查提供了巨大的潜力。