Hormone responsive tissue culture system and uses thereof

Abstract

The invention provides tissue culture system for primary cells (e.g. normal mammalian primary epithelial progenitors). This system includes: a) a serum-free, chemically defined cell culture media; and, b) methods for isolation and in vitro long-term propagation of primary cells (e.g. primary epithelial cells). Primary cells so isolated and cultured can be kept undifferentiated and proliferate for many weeks (>15 weeks) or population doubling (>35 PD) without senescence, or any detectable genetic alterations. Upon changing media / culture conditions, these cells can be induced to differentiate.The invention also provides methods to transform normal primary cells so cultured into “cancer stem cells.” The genetically defined cancer stem cell tumor model mimics the behavior of the disease closely, e.g., the cells are invasive, hormone responsive and metastatic when injected into mice. The tumor cells express genes that are specific to cancer stem cells identified in patient samples.

Description

RELATED APPLICATIONS[0001]This application claims the benefit of the filing date of U.S. provisional application 60 / 964,271, entitled “Hormone Responsive Tissue Culture System and Uses Thereof,” filed Aug. 10, 2007. This application is related to U.S. provisional application 60 / 569,005, entitled “Contribution of Target Cell Type to Epithelial Tumor Phenotypes,” filed May 7, 2004 and U.S. provisional application 60 / 630,934, entitled “Hormone Responsive Tissue Culture System and Uses Thereof,” filed Nov. 24, 2004. The entire teachings of the referenced applications are expressly incorporated herein by reference.GOVERNMENT SUPPORT[0002]The invention described herein was supported, in whole or in part, by Grant No. K08-CA-92013 from the National Cancer Institute (NCI). The U.S. Government has certain rights in this invention.BACKGROUND OF THE INVENTION[0003]Recent work comparing transformation and tumorigenicity of rodent and human cells has established significant differences between s...

AI Technical Summary

Benefits of technology

[0010]The medium of the present invention supports the growth of primary mammary epithelial progenitor cells that can give rise to a luminal epithelial phenotype and simultaneously suppresses the growth of other cell types. Thus, primary cells isolated using such a medium / system is essentially free of other cell types, especially those undesirable cell types, such as myoepithelial, stromal, and basal epithelial cell types.

[0053]In certain embodiments, the medium supports growth, proliferation, and / or differentiation of primary epithelial progenitor cells induced to express: (1) telomerase catalytic subunit, (2) a first polypeptide that functions in the same signaling pathway(s) as the SV40 large T antigen, and (3) a second polypeptide that functions in the same signaling pathway as the mutant H-ras oncogene product, for at least about 15 weeks or at least about 35 population doubling (PD) in vitro, without losing tumorigenicity.

Problems solved by technology

It has been difficult to retrospectively identify the precise cell type that gives rise to a particular tumor in clinical samples or rodent tumor models, since the normal cell from which the tumor arose is already transformed and no longer available in its original state.

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