J Cancer 2012; 3:196-206. doi:10.7150/jca.4484

Research Paper

Establishment of Human Ultra-Low Passage Colorectal Cancer Cell Lines Using Spheroids from Fresh Surgical Specimens Suitable for In Vitro and In Vivo Studies

Satyajit Ray1, Russell C. Langan1, John E. Mullinax1, Tomotake Koizumi1, Hong-Wu Xin1, Gordon W. Wiegand1, Andrew J. Anderson1, Alexander Stojadinovic2,3, Snorri Thorgeirsson4✉, Udo Rudloff1, Itzhak Avital1,5 ✉

1. Surgery Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD;
2. Department of Surgery, Division of Surgical Oncology, Walter Reed National Military Medical Center, Bethesda, MD;
3. Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, MD;
4. Laboratory of Experimental Carcinogenesis, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD;
5. Bon Secours Cancer Institute, Richmond VA, USA

Abstract

Colorectal cancer (CRC) holds the third highest incidence and cancer related mortality rate among men and women in the United States. Unfortunately, there has been little progression made in the treatment of this deadly disease once it has spread beyond the colon. It has been hypothesize that colon cancer stem cells are implicated in CRC carcinogenesis, metastasis, and therapeutic resistance. One of the difficulties in testing these hypotheses is the current use of established high-passage cancer cell lines. Long term, high-passage established cell lines have cells with stem like properties as they propagate almost indefinitely. These cells are thought to be different than the original cancer stem cells in fresh tumors. In order to investigate cancer stem cells, and molecularly profiling tumors with high fidelity to the original primary tumor, one needs to establish suitable primary ultra-low passage cell lines from fresh surgical specimens. Here we report the establishment of tumor initiating colon cancer ultra-low passage cell lines by a combination of gentle mechanical, enzymatic dissociation, spheroid formation, and followed by two generation xenografts from fresh tumors obtained at time of operation. Tumors generated were characterized by morphology, flow cytometry, immunofluorescence, and by gene expression. In the future, such a technology can be used to produce expeditiously enough material to test for mutations, genetic signatures and molecular subtyping readily available for clinical therapeutic decision making.

Keywords: Colon cancer, Fresh Tumor, Cell lines, Spheroids, Cancer Stem Cells.

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How to cite this article:
Ray S, Langan RC, Mullinax JE, Koizumi T, Xin HW, Wiegand GW, Anderson AJ, Stojadinovic A, Thorgeirsson S, Rudloff U, Avital I. Establishment of Human Ultra-Low Passage Colorectal Cancer Cell Lines Using Spheroids from Fresh Surgical Specimens Suitable for In Vitro and In Vivo Studies. J Cancer 2012; 3:196-206. doi:10.7150/jca.4484. Available from http://www.jcancer.org/v03p0196.htm