, Volume 70, Issue 2, pp 567–576 | Cite as

Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro

  • Katharina S. Mießler
  • Constanze Vitzthum
  • Alexander G. Markov
  • Salah AmashehEmail author
Original Article


Mammary gland epithelium is physiologically exposed to variations of hydrostatic pressure due to accumulation of milk and removal by suckling and mechanical milking. Integrity of the mammary gland epithelium primarily relies on the tight junction. To analyze pressure-induced effects on the tight junction, we established a modified Ussing chamber and tested the hypothesis if hydrostatic pressure on the basal side of the epithelium is able to affect barrier properties in a mammary epithelial cell model, in vitro. Therefore, a conventional Ussing chamber was modified by an additional tube system to apply hydrostatic pressure. Monolayers of the mammary epithelial cell line HC11 were mounted in the modified Ussing chambers and incubated with increasing basal hydrostatic pressure. Transepithelial resistance and short circuit current were recorded and compared to controls. Hydrostatic pressure was stably applied and incubation steps of 30 min were technically feasible, leading to a decrease of transepithelial resistance and an increase of short circuit current in all monolayers. In a series of experiments simulating the physiological exposure time by short intervals of 5 min, these electrophysiological findings were also observed, and monolayer integrity was not significantly perturbed as analyzed by fluorescence immunohistochemistry selectively staining tight junction proteins. Moreover, electrophysiology demonstrated reversibility of effects. In conclusion, the modified Ussing chamber is an adequate method to analyze the effects of hydrostatic pressure on epithelial cell monolayers, in vitro. Both, the reduction of transepithelial resistance and the increase of short circuit current may be interpreted as protective reactions.


Barrier function Tight junction Ussing chamber Hydrostatic pressure Mammary gland HC11 



We thank Martin Grunau, Katharina Söllig, Uwe Tietjen, and Susanne Trappe for excellent technical assistance. This work was supported by the Center for International Cooperation of the Freie Universität Berlin [Grant No. FSP2016-300], the Deutsche Forschungsgemeinschaft [Grant No. AM 141/11-1], and the Sonnenfeld-Stiftung [doctoral studies Grant to K.S. Mießler].

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10616_2017_130_MOESM1_ESM.pdf (376 kb)
Supplementary material 1 (PDF 375 kb)


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Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Katharina S. Mießler
    • 1
  • Constanze Vitzthum
    • 1
  • Alexander G. Markov
    • 2
  • Salah Amasheh
    • 1
    Email author
  1. 1.Institute of Veterinary PhysiologyFreie Universität BerlinBerlinGermany
  2. 2.Department of General PhysiologySt. Petersburg State UniversitySaint PetersburgRussia

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