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Novel Cell Culture System for Primary Culture of Epithelium from Breast, Ovarian and Endometrial Tissues

Detailed Technology Description

None

*Abstract

It would be most useful if epithelium tissue could be grown in vitro conveniently and would exhibit the morphological, physiological, and biochemical behavior characteristic of epithelial cells grown in vivo. These capabilities would allow investigation in vitro of both the development of epithelial cancer, in terms of what triggers such a cancer and how it proceeds, and how better to treat or prevent it. A method and a medium for culturing epithelial cells of both normal and malignant origin is provided. The method entails physically disaggregating tissue samples, placing the resulting fragments onto a surface comprised of basement membrane matrix components, and culturing the tissue in a medium containing preselected fetal and newborn calf sera and rat sera. Both primary explant cell cultures and cell lines, which are long-lived and particularly suitable for further study, are produced. The cultured primary explant cells undergo differentiation to form complex structures resembling those seen in vivo. It is therefore an object of the present invention to provide an improved culture method which is simple and inexpensive, and which results in cultured epithelial tissue that is long-lived and that produces secondary structures that resemble epithelial structures in vivo. As an example, such structures in breast tissue are duct-like, branching and luminal. The culture method includes less stringent and elaborate initial tissue disaggregation techniques, an improved culture medium for use in the initial establishment of the cultures, the presence of extracellular matrix components in the culture chamber, and an optimal density of living cells in the culture chamber. The method can be used to produce primary explant cell cultures, secondary or extended explant cell cultures, or cultures of immortal cell lines from epithelial tissue. The cultures obtained by this method survive for at least 8 weeks, as a primary culture. Alternatively, the cultures may be frozen for subsequent use.

*Principal Investigator

Name: Jean Latimer

Department: Med-OB-Gyn & Reproductive Science

Country/Region

USA