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Mießler KS, Vitzthum C, Markov AG, et al. Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro. Cytotechnology. 2017;70(2):567-576doi: 10.1007/s10616-017-0130-3.
Mießler, K. S., Vitzthum, C., Markov, A. G., & Amasheh, S. (2018). Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro. Cytotechnology, 70(2), 567-576. https://doi.org/10.1007/s10616-017-0130-3
Mießler, Katharina S, et al. "Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro." Cytotechnology vol. 70,2 (2018): 567-576. doi: https://doi.org/10.1007/s10616-017-0130-3
Mießler KS, Vitzthum C, Markov AG, Amasheh S. Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro. Cytotechnology. 2018 Apr;70(2):567-576. doi: 10.1007/s10616-017-0130-3. Epub 2017 Aug 29. PMID: 28852895; PMCID: PMC5851952.
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Cytotechnology. 2018 Apr;70(2):567-576. doi: 10.1007/s10616-017-0130-3. Epub 2017 Aug 29.

Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro.

Cytotechnology

Katharina S Mießler, Constanze Vitzthum, Alexander G Markov, Salah Amasheh

Affiliations

  1. Institute of Veterinary Physiology, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany.
  2. Department of General Physiology, St. Petersburg State University, Universitetskaya nab. 7/9, Saint Petersburg, Russia, 199034.
  3. Institute of Veterinary Physiology, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany. [email protected].

PMID: 28852895 PMCID: PMC5851952 DOI: 10.1007/s10616-017-0130-3

Abstract

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.

Keywords: Barrier function; HC11; Hydrostatic pressure; Mammary gland; Tight junction; Ussing chamber

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