Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro. doi: 10.1007/s10616-017-0130-3 Q2

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

Research output

2 Citations (Scopus)

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
Original languageEnglish
Article number10.1007/s10616-017-0130-3
Pages (from-to)567-576
Number of pages10
JournalCytotechnology
Volume70
Issue number2
Early online date29 Aug 2017
DOIs
Publication statusPublished - 1 Apr 2018

Fingerprint

breasts
Monolayers
Breast
epithelial cells
Epithelial Cells
Hydrostatic pressure
Pressure
Short circuit currents
Hydrostatic Pressure
Epithelium
Electrophysiology
Tight Junction Proteins
Pressure effects
tight junctions
epithelium
Tight Junctions
Human Mammary Glands
Fluorescence
Cells
Proteins

Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Mießler, Katharina S. ; Vitzthum, Constanze ; Markov, Alexander G. ; Amasheh, Salah. / Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro. doi: 10.1007/s10616-017-0130-3 Q2. In: Cytotechnology. 2018 ; Vol. 70, No. 2. pp. 567-576.
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abstract = "Mammary gland epithelium is physiologicallyexposed to variations of hydrostatic pressuredue to accumulation of milk and removal by sucklingand mechanical milking. Integrity of the mammarygland epithelium primarily relies on the tight junction.To analyze pressure-induced effects on the tightjunction, we established a modified Ussing chamberand tested the hypothesis if hydrostatic pressure on thebasal side of the epithelium is able to affect barrierproperties in a mammary epithelial cell model,in vitro. Therefore, a conventional Ussing chamberwas modified by an additional tube system to applyhydrostatic pressure. Monolayers of the mammaryepithelial cell line HC11 were mounted in the modifiedUssing chambers and incubated with increasing basalhydrostatic pressure. Transepithelial resistance andshort circuit current were recorded and compared tocontrols. Hydrostatic pressure was stably applied andincubation steps of 30 min were technically feasible,leading to a decrease of transepithelial resistance andan increase of short circuit current in all monolayers.In a series of experiments simulating the physiologicalexposure time by short intervals of 5 min, theseelectrophysiological findings were also observed, andmonolayer integrity was not significantly perturbed asanalyzed by fluorescence immunohistochemistryselectively staining tight junction proteins. Moreover,electrophysiology demonstrated reversibility ofeffects. In conclusion, the modified Ussing chamberis an adequate method to analyze the effects ofhydrostatic pressure on epithelial cell monolayers,in vitro. Both, the reduction of transepithelial resistanceand the increase of short circuit current may beinterpreted as protective reactions",
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Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro. doi: 10.1007/s10616-017-0130-3 Q2. / Mießler, Katharina S. ; Vitzthum, Constanze; Markov, Alexander G.; Amasheh, Salah.

In: Cytotechnology, Vol. 70, No. 2, 10.1007/s10616-017-0130-3, 01.04.2018, p. 567-576.

Research output

TY - JOUR

T1 - Basolateral pressure challenges mammary epithelial cell monolayer integrity, in vitro. doi: 10.1007/s10616-017-0130-3 Q2

AU - Mießler, Katharina S.

AU - Vitzthum, Constanze

AU - Markov, Alexander G.

AU - Amasheh, Salah

PY - 2018/4/1

Y1 - 2018/4/1

N2 - Mammary gland epithelium is physiologicallyexposed to variations of hydrostatic pressuredue to accumulation of milk and removal by sucklingand mechanical milking. Integrity of the mammarygland epithelium primarily relies on the tight junction.To analyze pressure-induced effects on the tightjunction, we established a modified Ussing chamberand tested the hypothesis if hydrostatic pressure on thebasal side of the epithelium is able to affect barrierproperties in a mammary epithelial cell model,in vitro. Therefore, a conventional Ussing chamberwas modified by an additional tube system to applyhydrostatic pressure. Monolayers of the mammaryepithelial cell line HC11 were mounted in the modifiedUssing chambers and incubated with increasing basalhydrostatic pressure. Transepithelial resistance andshort circuit current were recorded and compared tocontrols. Hydrostatic pressure was stably applied andincubation steps of 30 min were technically feasible,leading to a decrease of transepithelial resistance andan increase of short circuit current in all monolayers.In a series of experiments simulating the physiologicalexposure time by short intervals of 5 min, theseelectrophysiological findings were also observed, andmonolayer integrity was not significantly perturbed asanalyzed by fluorescence immunohistochemistryselectively staining tight junction proteins. Moreover,electrophysiology demonstrated reversibility ofeffects. In conclusion, the modified Ussing chamberis an adequate method to analyze the effects ofhydrostatic pressure on epithelial cell monolayers,in vitro. Both, the reduction of transepithelial resistanceand the increase of short circuit current may beinterpreted as protective reactions

AB - Mammary gland epithelium is physiologicallyexposed to variations of hydrostatic pressuredue to accumulation of milk and removal by sucklingand mechanical milking. Integrity of the mammarygland epithelium primarily relies on the tight junction.To analyze pressure-induced effects on the tightjunction, we established a modified Ussing chamberand tested the hypothesis if hydrostatic pressure on thebasal side of the epithelium is able to affect barrierproperties in a mammary epithelial cell model,in vitro. Therefore, a conventional Ussing chamberwas modified by an additional tube system to applyhydrostatic pressure. Monolayers of the mammaryepithelial cell line HC11 were mounted in the modifiedUssing chambers and incubated with increasing basalhydrostatic pressure. Transepithelial resistance andshort circuit current were recorded and compared tocontrols. Hydrostatic pressure was stably applied andincubation steps of 30 min were technically feasible,leading to a decrease of transepithelial resistance andan increase of short circuit current in all monolayers.In a series of experiments simulating the physiologicalexposure time by short intervals of 5 min, theseelectrophysiological findings were also observed, andmonolayer integrity was not significantly perturbed asanalyzed by fluorescence immunohistochemistryselectively staining tight junction proteins. Moreover,electrophysiology demonstrated reversibility ofeffects. In conclusion, the modified Ussing chamberis an adequate method to analyze the effects ofhydrostatic pressure on epithelial cell monolayers,in vitro. Both, the reduction of transepithelial resistanceand the increase of short circuit current may beinterpreted as protective reactions

KW - Barrier function; HC11; Hydrostatic pressure; Mammary gland; Tight junction; Ussing chamber

KW - Barrier function Tight junction Ussing chamber Hydrostatic pressure Mammary gland HC11

U2 - 10.1007/s10616-017-0130-3

DO - 10.1007/s10616-017-0130-3

M3 - Article

VL - 70

SP - 567

EP - 576

JO - Cytotechnology

JF - Cytotechnology

SN - 0920-9069

IS - 2

M1 - 10.1007/s10616-017-0130-3

ER -