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Collagen Gel Method for the Primary Culture of Mouse Mammary Epithelium

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Methods in Mammary Gland Biology and Breast Cancer Research

Abstract

A method for culturing mouse mammary epithelial cells (MECs) within Type I collagen gels is described. This method is permissive for multifold growth, “ductlike” morphogenesis, and hormonal responsiveness. Subcutaneous mouse mammary glands are harvested, digested with collagenase, and epithelial cells obtained after Percoll gradient centrifugation. MECs are then mixed with neutralized, isosmotic collagen, which is pipetted into culture dishes and overlaid with culture medium after gel formation. Preparation of all serum-free medium and stocks required for the procedure and termination of cultures are described, and comments concerning critical procedural steps are given.

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Abbreviations

(MECs):

mammary epithelial cells

(M199):

medium 199

(pen/strep):

penicillin/streptomycin

(Amp B):

Amphotericin B

(HEPES):

N-(2-hydroxyethyl)piperazine-N’-(2-ethanesulfonic acid)

(DMEM):

Dulbecco’s modified Eagle’s medium

(BSA):

bovine serum albumin

(HBSS):

Hanks’ balanced salt solution

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Author information

Authors and Affiliations

  1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas, 66160, USA

    Walter Imagawa

  2. Department of Molecular and Cell Biology, Cancer Research Laboratory, University of California, Berkeley, California, 94720, USA

    Jason Yang, Raphael C. Guzman & Satyabrata Nandi

Authors
  1. Walter Imagawa
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  2. Jason Yang
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  3. Raphael C. Guzman
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  4. Satyabrata Nandi
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Editor information

Editors and Affiliations

  1. Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, New York, 14263, USA

    Margot M. Ip

  2. Roswell Park Cancer Research Institute, Buffalo, New York, USA

    Bonnie B. Asch

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© 2000 Springer Science+Business Media New York

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Imagawa, W., Yang, J., Guzman, R.C., Nandi, S. (2000). Collagen Gel Method for the Primary Culture of Mouse Mammary Epithelium. In: Ip, M.M., Asch, B.B. (eds) Methods in Mammary Gland Biology and Breast Cancer Research. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-4295-7_11

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  • DOI: https://doi.org/10.1007/978-1-4615-4295-7_11

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-6927-1

  • Online ISBN: 978-1-4615-4295-7

  • eBook Packages: Springer Book Archive

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