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BMC Cancer. 2012; 12: 10.
Published online Jan 10, 2012. doi:  10.1186/1471-2407-12-10
PMCID: PMC3277457
"A novel in vivo model for the study of human breast cancer metastasis using primary breast tumor-initiating cells from patient biopsies"
Carolyn G Marsden,1 Mary Jo Wright,2 Latonya Carrier,3 Krzysztof Moroz,4 Radhika Pochampally,5 and Brian G Rowancorresponding author6
1Department of Structural and Cellular Biology, Tulane University Health Sciences Center, The Louisiana Cancer Research Consortium, New Orleans, LA 70112, USA
2Department of Surgery, Tulane University School of Medicine, The Louisiana Cancer Research Consortium, New Orleans, LA 70112, USA
3Department of Structural and Cellular Biology, Tulane University Health Sciences Center, New Orleans, LA 70112, USA
4Section of Surgical Pathology & Cytopathology, Tulane University School of Medicine, Louisiana Cancer Research Consortium, New Orleans, LA 70112, USA
5Department of Pharmacology, Tulane University Health Sciences Center, Center for Gene Therapy, Louisiana Cancer Research Consortium, New Orleans, LA, 70,112, USA
6Department of Structural and Cellular Biology, Tulane University Health Sciences Center, Louisiana Cancer Research Consortium, Center for Gene Therapy, New Orleans, LA 70112, USA
corresponding authorCorresponding author.
Carolyn G Marsden: cmarsden/at/tulane.edu; Mary Jo Wright: mjwright/at/tulane.edu; Latonya Carrier: lcarrie/at/tulane.edu; Krzysztof Moroz: kmoroz/at/tulane.edu; Radhika Pochampally: rpocham/at/tulane.edu; Brian G Rowan: browan/at/tulane.edu
Received October 12, 2011; Accepted January 10, 2012.
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 α (ERα)/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 mammary fat pad of NUDE mice. At 3 months post-injection, micrometastases to the lung, liver, kidneys, brain and femur were detected by measuring content of human chromosome 17. Visible macrometastases were detected in the lung, liver and kidneys by 6 months post-injection. Primary tumors variably expressed cytokeratins, Her2/neu, cytoplasmic E-cadherin, nuclear β catenin and fibronectin but were negative for ERα and vimentin. In lung and liver metastases, variable redistribution of E-cadherin and β catenin to the membrane of tumor cells was observed. ERα was re-expressed in lung metastatic cells in two of five samples.
Conclusions
Tumorspheres isolated under defined culture conditions from patient core biopsies were tumorigenic when transplanted into the mammary fat pad of NUDE mice, and metastasized to multiple mouse organs. Micrometastases in mouse organs demonstrated a dormancy period prior to outgrowth of macrometastases. The development of macrometastases with organ-specific phenotypic distinctions provides a superior model for the investigation of organ-specific effects on metastatic cancer cell survival and growth.
Keywords: Primary breast tumor-initiating cells, Metastasis, Dormancy, EMT
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