title: Ð ÐµÐ³Ð¸ÑÑÑÐ°ÑÐ¸Ñ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð¸Ð¾Ð½Ð½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð² Ð¼ÐµÐ·ÐµÐ½Ñ Ð¸Ð¼Ð½ÑÑ ÑÑÐ²Ð¾Ð»Ð¾Ð²ÑÑ ÐºÐ»ÐµÑÐºÐ°Ñ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ° Ð¿ÑÐ¸ 3-D ÐºÑÐ»ÑÑÐ¸Ð²Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ð¸
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¼Ð°Ð³Ð¸ÑÑÑÐ°
titleType: Subtitle
title: Ion channel recording in 3D culture of mesenchymal stem cells
Creators: name: Ð¥Ð°Ð¹ÑÑÐ»Ð»Ð¸Ð½Ð°, ÐÑÐ»ÐµÐ¹ÑÐ°
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2022
Subjects: ÐÐ¸Ð¾ÑÐ¸Ð·Ð¸ÐºÐ°
ÐÐ¾Ð½Ñ
ÐÐ»ÐµÑÐºÐ° (Ð±Ð¸Ð¾Ð».)
Ð¼ÐµÑÐ¾Ð´ Ð²Ð¸ÑÑÑÐµÐ¹ ÐºÐ°Ð¿Ð»Ð¸
Ð¸Ð¾Ð½Ð½ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ
Ð¼ÐµÑÐ¾Ð´ Ð»Ð¾ÐºÐ°Ð»ÑÐ½Ð¾Ð¹ ÑÐ¸ÐºÑÐ°ÑÐ¸Ð¸ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»Ð° (Ð¿Ð°ÑÑ-ÐºÐ»Ð°Ð¼Ð¿)
ÐºÐ°Ð»ÑÑÐ¸ÐµÐ²Ð°Ñ ÑÐ¸Ð³Ð½Ð°Ð»Ð¸Ð·Ð°ÑÐ¸Ñ
ÐºÐ°Ð»ÑÑÐ¸Ð¹-Ð·Ð°Ð²Ð¸ÑÐ¸Ð¼ÑÐµ ÐºÐ°Ð»Ð¸ÐµÐ²ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ
Ð¼ÐµÑ Ð°Ð½Ð¾ÑÑÐ²ÑÑÐ²Ð¸ÑÐµÐ»ÑÐ½ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ
hanging drop method
ion channels
patch clamp
calcium signaling
calcium-dependent potassium channels
mechanosensitive channels

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐºÐµ Ð¸ Ð°Ð¿ÑÐ¾Ð±Ð°ÑÐ¸Ð¸ Ð¼ÐµÑÐ¾Ð´Ð° Ð»Ð¾ÐºÐ°Ð»ÑÐ½Ð¾Ð¹ ÑÐ¸ÐºÑÐ°ÑÐ¸Ð¸ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»Ð° (Ð¿Ð°ÑÑ-ÐºÐ»Ð°Ð¼Ð¿) Ð´Ð»Ñ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð½Ð°ÑÐ¸Ð²Ð½ÑÑ Ð¸Ð¾Ð½Ð½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð² Ð¿Ð»Ð°Ð·Ð¼Ð°ÑÐ¸ÑÐµÑÐºÐ¾Ð¹ Ð¼ÐµÐ¼Ð±ÑÐ°Ð½Ðµ Ð¼ÐµÐ·ÐµÐ½ÑÐ¸Ð¼Ð½ÑÑ ÑÑÐ²Ð¾Ð»Ð¾Ð²ÑÑ ÐºÐ»ÐµÑÐ¾Ðº ÑÐ½Ð´Ð¾Ð¼ÐµÑÑÐ¸Ñ (ÑÐÐ¡Ð) ÑÐµÐ»Ð¾Ð²ÐµÐºÐ° Ð¿ÑÐ¸ 3D ÐºÑÐ»ÑÑÐ¸Ð²Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ð¸.Â Ð¢ÐµÐ¼Ð°ÑÐ¸ÐºÐ° Ð½Ð°ÑÑÐ½Ð¾Ð³Ð¾ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ ÑÐ²Ð»ÑÐµÑÑÑ Ð²Ð°Ð¶Ð½Ð¾Ð¹ Ð² ÑÐ°Ð¼ÐºÐ°Ñ Ð²Ð¾Ð¿ÑÐ¾ÑÐ° Ð²ÑÑÐ²Ð»ÐµÐ½Ð¸Ñ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾Ð¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð¸Ð¾Ð½Ð½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð¸ Ð¸Ñ ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ Ð² ÑÐ¸Ð·Ð¸Ð¾Ð»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¸Ñ ÑÐµÐ°ÐºÑÐ¸ÑÑ ÑÑÐµÑÐ¼ÐµÑÐ½ÑÑ ÐºÐ»ÐµÑÐ¾ÑÐ½ÑÑ ÐºÑÐ»ÑÑÑÑ. ÐÑÐ½Ð¾Ð²Ð½Ð¾Ð¹ ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½ÑÐ°Ð»ÑÐ½ÑÐ¹ Ð¿Ð¾Ð´ÑÐ¾Ð´ - Ð¼ÐµÑÐ¾Ð´ Ð¿Ð°ÑÑ-ÐºÐ»Ð°Ð¼Ð¿ Ð² ÐºÐ¾Ð½ÑÐ¸Ð³ÑÑÐ°ÑÐ¸Ð¸ cell-attached, Ð¿Ð¾Ð·Ð²Ð¾Ð»ÑÑÑÐ¸Ð¹ Ð·Ð°Ð¿Ð¸ÑÑÐ²Ð°ÑÑ Ð¸Ð¾Ð½Ð½ÑÐµ ÑÐ¾ÐºÐ¸ Ð¾Ð´Ð¸Ð½Ð¾ÑÐ½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð² Ð¸Ð·Ð¾Ð»Ð¸ÑÐ¾Ð²Ð°Ð½Ð½Ð¾Ð¼ ÑÑÐ°ÑÑÐºÐµ Ð¿Ð»Ð°Ð·Ð¼Ð°ÑÐ¸ÑÐµÑÐºÐ¾Ð¹ Ð¼ÐµÐ¼Ð±ÑÐ°Ð½Ñ ÐºÐ»ÐµÑÐ¾Ðº. ÐÐ»Ñ ÑÐµÐ³Ð¸ÑÑÑÐ°ÑÐ¸Ð¸ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð¸Ð¾Ð½Ð½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² ÑÑÐµÑÐ¼ÐµÑÐ½ÑÑ ÐºÐ»ÐµÑÐ¾ÑÐ½ÑÑ Ð°Ð³ÑÐµÐ³Ð°ÑÐ¾Ð² Ð½Ð°Ð¼Ð¸ Ð±ÑÐ» ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐ°Ð½ Ð½Ð¾Ð²ÑÐ¹ Ð¿Ð¾Ð´ÑÐ¾Ð´ Ð´Ð»Ñ Ð¾Ð±ÑÐ°Ð·Ð¾Ð²Ð°Ð½Ð¸Ñ ÑÑÐ°Ð±Ð¸Ð»ÑÐ½Ð¾Ð³Ð¾ ÑÐ²ÐµÑÑÐ¿Ð»Ð¾ÑÐ½Ð¾Ð³Ð¾ ÐºÐ¾Ð½ÑÐ°ÐºÑÐ° Ð¼ÐµÐ¶Ð´Ñ ÑÐµÐ³Ð¸ÑÑÑÐ¸ÑÑÑÑÐµÐ¹ Ð¿Ð¸Ð¿ÐµÑÐºÐ¾Ð¹ Ð¸ ÑÑÐ°Ð³Ð¼ÐµÐ½ÑÐ¾Ð¼ Ð¼ÐµÐ¼Ð±ÑÐ°Ð½Ñ ÐºÐ»ÐµÑÐ¾Ðº, Ð°Ð³ÑÐµÐ³Ð¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ Ð² ÑÑÐµÑÐ¾Ð¸Ð´Ñ. Ð¡ Ð¿Ð¾Ð¼Ð¾ÑÑÑ Ð´Ð°Ð½Ð½Ð¾Ð³Ð¾ Ð¿ÑÐ¾ÑÐ¾ÐºÐ¾Ð»Ð° Ð½Ð°Ð¼Ð¸ Ð²Ð¿ÐµÑÐ²ÑÐµ Ð±ÑÐ»Ð° Ð·Ð°ÑÐµÐ³Ð¸ÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð° Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ð¼ÐµÑÐ°Ð½Ð¾ÑÑÐ²ÑÑÐ²Ð¸ÑÐµÐ»ÑÐ½ÑÑ Ð¸Ð¾Ð½Ð½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² (ÐÐ§) Ð² ÐºÐ»ÐµÑÐºÐ°Ñ ÑÐÐ¡Ð, ÑÐ¾Ð±ÑÐ°Ð½Ð½ÑÑ Ð² ÑÑÐµÑÐ¾Ð¸Ð´Ñ. ÐÐ¾Ð»ÐµÐµ ÑÐ¾Ð³Ð¾, Ð¼Ñ Ð½Ð°Ð±Ð»ÑÐ´Ð°Ð»Ð¸ Ð¸Ñ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾Ðµ Ð²Ð·Ð°Ð¸Ð¼Ð¾Ð´ÐµÐ¹ÑÑÐ²Ð¸Ðµ Ñ ÐºÐ°Ð»Ð¸ÐµÐ²ÑÐ¼Ð¸ ÐºÐ°Ð½Ð°Ð»Ð°Ð¼Ð¸ Ð±Ð¾Ð»ÑÑÐ¾Ð¹ Ð¿ÑÐ¾Ð²Ð¾Ð´Ð¸Ð¼Ð¾ÑÑÐ¸ (BK). Ð¡ Ð¿Ð¾Ð¼Ð¾ÑÑÑ Ð¸Ð¼Ð¼ÑÐ½Ð¾ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾Ð³Ð¾ Ð¾ÐºÑÐ°ÑÐ¸Ð²Ð°Ð½Ð¸Ñ Ð±ÑÐ»Ð° Ð¿Ð¾ÐºÐ°Ð·Ð°Ð½Ð° ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ñ BK ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð² Ð¼ÐµÐ¼Ð±ÑÐ°Ð½Ðµ ÐºÐ»ÐµÑÐ¾Ðº ÑÐÐ¡Ð, Ð°Ð³ÑÐµÐ³Ð¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ Ð² ÑÑÐµÑÐ¾Ð¸Ð´Ñ. Ð¢Ð°ÐºÐ¶Ðµ Ð¼Ñ Ð½Ð°Ð±Ð»ÑÐ´Ð°Ð»Ð¸ Ð´Ð¾ÑÑÐ¾Ð²ÐµÑÐ½Ð¾Ðµ ÑÐ½Ð¸Ð¶ÐµÐ½Ð¸Ðµ ÑÐ°ÑÑÐ¾ÑÑ Ð°ÐºÑÐ¸Ð²Ð°ÑÐ¸Ð¸ ÐÐ§ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð² ÑÑÐµÑÐ¾Ð¸Ð´Ð°Ñ, ÑÑÐ¾ Ð¼Ð¾Ð¶ÐµÑ ÑÐ²Ð¸Ð´ÐµÑÐµÐ»ÑÑÑÐ²Ð¾Ð²Ð°ÑÑ Ð¾ ÑÐ°Ð·Ð»Ð¸ÑÐ¸ÑÑ Ð² ÑÑÐ¾Ð²Ð½Ðµ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾Ð¹ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð² 3D ÐºÑÐ»ÑÑÑÑÐµ Ð¿Ð¾ ÑÑÐ°Ð²Ð½ÐµÐ½Ð¸Ñ Ñ 2D ÐºÑÐ»ÑÑÑÑÐ¾Ð¹ ÑÐÐ¡Ð.
description: The present work is devoted to development and application of the specific variant of the patch-clamp method for the recording of the activity of native ion channels in 3D spheroids formed from human endometrial mesenchymal stem cells (eMSCs). The research area is important in terms of registration of the functional activity of ion channels and their regulation in the physiological reactions in 3D cell cultures. The main experimental approach is the patch-clamp method in the cell-attached configuration (registration of single ion channels). To register the ion channel activity in three-dimensional cell aggregates, we developed a new approach for the formation of the stable contact between recording pipette and plasma membrane fragment of the cells aggregated into spheroids. Using the developed approach, we have recorded the activity of mechanosensitive ion channels eMSCs assembled in spheroids. Moreover, we observed their functional interplay with potassium channels of big conductance (BK) in the plasma membrane. Using immunofluorescent staining the BK expression in the membrane of eMSCs was shown. Also, we observed a significant decrease in the frequency of mechanosensitive channel activation in spheroids that may indicate the differences in the level of functional expression of channels in 3D culture comparing to 2D culture of eMSCs.