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Primary culture and immortalization of human fallopian tube secretory epithelial cells

Nature Protocols volume 7, pages 1755–1764 (2012)Cite this article

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Abstract

Primary human fallopian tube secretory epithelial cell (FTSEC) cultures are useful for studying normal fallopian tube epithelial biology, as well as for developing models of fallopian tube disease, such as cancer. Because of the limited ability of primary human FTSECs to proliferate in vitro, it is necessary to immortalize them in order to establish a cell line that is suitable for long-term culture and large-scale in vitro experimentation. This protocol describes the isolation of FTSECs from human fallopian tube tissue, conditions for primary FTSEC culture and techniques for establishing immortal FTSEC lines. The entire process, from primary cell isolation to establishment of an immortal cell line, may take up to 2 months. Once established, immortal FTSECs can typically be maintained for at least 30 passages.

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Figure 1: Methods for culturing primary human fallopian tube epithelial cells.
Figure 2: Fallopian tube tissue processing.
Figure 3: Seeding and culture of dissociated FTSECs on collagen-coated plasticware.
Figure 4: Emergence of FTSECs colonies under selective pressure.
Figure 5: Expansion of immortal FTSECs.
Figure 6: Validation of immortal FTSECs.

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Acknowledgements

Special thanks to the faculty and staff of the Brigham and Women's Hospital Department of Pathology for allocation of tissues. This work was supported by a Canadian Institutes of Health Research Fellowship (A.M.K.), a Kaleidoscope of Hope Young Investigator Research Grant (A.M.K.), US National Institutes of Health grant no. P50 CA105009 (SPORE), the Ovarian Cancer Research Fund (R.D.), the Robert and Debra First Fund (R.D.), the Randi and Joel Cutler Ovarian Cancer Research Fund (R.D.), the Mary Kay Foundation (R.D.), the Sandy Rollman Ovarian Cancer Foundation (R.D.) and the Susan Smith Center for Women's Cancers at the Dana-Farber Cancer Institute (R.D.).

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Authors and Affiliations

  1. Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, USA

    Alison M Karst & Ronny Drapkin

  2. Department of Pathology, Division of Women's and Perinatal Pathology, Brigham and Women's Hospital, Boston, Massachusetts, USA

    Ronny Drapkin

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  1. Alison M Karst
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Contributions

A.M.K. developed the experimental protocol, performed the experiments and wrote the manuscript. R.D. contributed to the conceptual design of the protocol and edited the manuscript.

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Correspondence to Ronny Drapkin.

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The authors declare no competing financial interests.

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Karst, A., Drapkin, R. Primary culture and immortalization of human fallopian tube secretory epithelial cells. Nat Protoc 7, 1755–1764 (2012). https://doi.org/10.1038/nprot.2012.097

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