Dani Sergio U, Espindola Rachel
AADM Gene Therapy Center, Av. Bandeirantes, 2419, 14030-670 Ribeirão Preto, SP, Brazil.
Genet Mol Res. 2002 Jun 30;1(2):167-75.
We developed a model system for testing gene vectors, based on the growth of murine tumors on the chorioallantoic membrane (CAM) of embryonic chickens. The ability of selected murine cells to grow on the CAM was rated according to the following criteria: i) formation of tumor masses; ii) metastasis formation; iii) reproducibility; iv) yield, indicated as the number of embryos surviving to assessment time with visible tumors on the CAM; v) maintainability of the cell, both in the original host and the embryonic chick, or 'shuttle maintainability'; vi) detection by the naked eye, and vii) cost/benefit relation. The murine melanoma cell lineage, B16F10, which efficiently forms distinct, pigmented tumor masses and metastases on the CAM, performed better in this model than the murine B61 cell line. In vitro transduction of B16F10 cells with a recombinant adenovirus carrying a construct of the E. coli LacZ gene followed by inoculation onto the CAM resulted in beta-galactosidase expression in the tumor mass growing on the CAM. This model is potentially applicable to preclinical evaluation of gene vectors, especially for gene therapy of cancer.
我们基于胚胎鸡绒毛尿囊膜(CAM)上小鼠肿瘤的生长情况,开发了一种用于测试基因载体的模型系统。根据以下标准对所选小鼠细胞在CAM上生长的能力进行评估:i)肿瘤块的形成;ii)转移灶的形成;iii)可重复性;iv)产量,以存活至评估时间且CAM上有可见肿瘤的胚胎数量表示;v)细胞在原始宿主和胚胎鸡中的维持能力,即“穿梭维持能力”;vi)肉眼可检测性;vii)成本效益关系。小鼠黑色素瘤细胞系B16F10在CAM上能高效形成明显的色素沉着肿瘤块和转移灶,在该模型中的表现优于小鼠B61细胞系。用携带大肠杆菌LacZ基因构建体的重组腺病毒对B16F10细胞进行体外转导,然后接种到CAM上,结果在CAM上生长的肿瘤块中出现了β-半乳糖苷酶表达。该模型可能适用于基因载体的临床前评估,特别是用于癌症的基因治疗。