Abstract
The success rate of establishing human cancer cell lines is not satisfactory and the established cell lines often do not preserve the molecular and histological features of the original tissues. In this study, we developed a novel culture method which can support proliferation of almost all primary epithelial ovarian cancer cells, as well as primary normal human oviductal epithelial cells. Cancer cells from fresh or frozen specimens were enriched by the anti-EpCAM antibody-conjugated magnetic beads, plated on Matrigel-coated plate and cultivated under the optimized culture conditions. Seventeen newly established ovarian cancer cell lines, which included all four major histotypes of ovarian cancer, were confirmed to express histotype-specific markers in vitro. Some of the cell lines from all the four histotypes, except mucinous type, generated tumors in immune-deficient mice and the xenograft tumor tissues recapitulated the corresponding original tissues faithfully. Furthermore, with poorly tumorigenic cell lines including mucinous type, we developed a novel xenograft model which could reconstruct the original tissue architecture through forced expression of a set of oncogenes followed by its silencing. With combination of the novel culture method and cell-derived xenograft system, virtually every epithelial ovarian cancer can be reconstituted in mice in a timely fashion.
Highlights
Cancer cell growth cannot be controlled inside the body, it is often difficult to culture in vitro and the success rate is poor in most cases [1,2]
HOV11T was established from an ovarian tumor which was diagnosed as adenocarcinoma NOS, it could be classified as high-grade serous carcinomas (HGSC) since HOV11T cells showed accumulation of TP53 and p16 and WT positivity (Figure 1)
We have established 17 ovarian tumor cell lines belonging to all four major histotypes including seven clear cell (CC) type, two EM type, three HGSC, one possible HGSC and three MC type lines, as well as a sarcomatous line derived from a MC tumor with sarcomatous components
Summary
Cancer cell growth cannot be controlled inside the body, it is often difficult to culture in vitro and the success rate is poor in most cases [1,2]. The reliability of commonly used cancer cell lines has recently been considered problematic. 47 ovarian cancer cell lines, some of the cell lines did not resemble cognate tumor profiles at all [4]. It is reported that many cell lines have acquired mutations not present in the original tumor. Most popular cancer cell line, HeLa, can continuously acquire novel mutations during passages, indicating strong selective pressure for cancer cells under ordinary culture conditions [5]
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