a novel in vivo model for the study of human breast cancer metastasis using primary breast tumor-initiating cells from patient biopsies一种新颖的体内模型研究人类乳腺癌转移利用患者的乳腺肿瘤起源细胞活检.pdfVIP

Marsden et al. BMC Cancer 2012, 12:10 /1471-2407/12/10 RESEARCH ARTICLE Open Access “A novel in vivo model for the study of human breast cancer metastasis using primary breast tumor-initiating cells from patient biopsies” 1 2 3 4 5 Carolyn G Marsden , Mary Jo Wright , Latonya Carrier , Krzysztof Moroz , Radhika Pochampally and Brian G Rowan6* Abstract Background: The study of breast cancer metastasis depends on the use of established breast cancer cell lines that do not accurately represent the heterogeneity and complexity of human breast tumors. A tumor model was developed using primary breast tumor-initiating cells isolated from patient core biopsies that would more accurately reflect human breast cancer metastasis. Methods: Tumorspheres were isolated under serum-free culture conditions from core biopsies collected from five patients with clinical diagnosis of invasive ductal carcinoma (IDC). Isolated tumorspheres were transplanted into the mammary fat pad of NUDE mice to establish tumorigenicity in vivo. Tumors and metastatic lesions were analyzed by hematoxylin and eosin (H+E) staining and immunohistochemistry (IHC). Results: Tumorspheres were successfully isolated from all patient core biopsies, independent of the estrogen receptor a (ERa)/progesterone receptor (PR)/Her2/neu status or tumor grade. Each tumorsphere was estimated to contain 50-100 cells. Transplantation of 50 tumorspheres (1-5 × 103 cells) in combination with Matrigel into the mammary fat pad of NUDE mice resulted in small, palpable tumors that were sustained up to 12 months post- injection. Tumors were serially transplanted three times by re-isolation of tumorspheres from the tumors and injection into the m