title: ÐÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ðµ ÑÐ¾Ð»Ð¸ ÑÐµÐ´ÑÐºÑÐ¸Ð²Ð½Ð¾Ð³Ð¾ ÑÑÑÐµÑÑÐ°, Ð²ÑÐ·Ð²Ð°Ð½Ð½Ð¾Ð³Ð¾ Ð¸Ð½Ð°ÐºÑÐ¸Ð²Ð°ÑÐ¸ÐµÐ¹ Ð´ÑÑ Ð°ÑÐµÐ»ÑÐ½Ð¾Ð¹ ÑÐµÐ¿Ð¸ Ð¼Ð¸ÑÐ¾Ñ Ð¾Ð½Ð´ÑÐ¸Ð¸, Ð² ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ Ð¿ÑÐ¾Ð»Ð¸ÑÐµÑÐ°ÑÐ¸Ð¸ ÑÐ¼Ð±ÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½ÑÑ ÑÑÐ²Ð¾Ð»Ð¾Ð²ÑÑ ÐºÐ»ÐµÑÐ¾Ðº
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Investigation of the role of reductive stress caused by inactivation of the mitochondrial respiratory chain in the regulation of embryonic stem cell proliferation
Creators: name: ÐÐ¸ÑÐ°ÑÐµÐ², ÐÐ³Ð¾ÑÑ
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: 2024
Subjects: ÑÑÐ²Ð¾Ð»Ð¾Ð²ÑÐµ ÐºÐ»ÐµÑÐºÐ¸
ÑÐµÐ´ÑÐºÑÐ¸Ð²Ð½ÑÐ¹ ÑÑÑÐµÑÑ
ÑÐ¼Ð±ÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½ÑÐµ ÑÑÐ²Ð¾Ð»Ð¾Ð²ÑÐµ ÐºÐ»ÐµÑÐºÐ¸
Ð¼Ð¸ÑÐ¾Ñ Ð¾Ð½Ð´ÑÐ¸Ð¸
Ð´ÐµÐ³Ð¸Ð´ÑÐ¾Ð³ÐµÐ½Ð°Ð·Ð½ÑÐ¹ ÐºÐ¾Ð¼Ð¿Ð»ÐµÐºÑ
stem cells
reductive stress
embryonic stem cells
mitochondria
dehydrogenase

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ ÑÐ¾Ð»Ð¸ ÑÐµÐ´ÑÐºÑÐ¸Ð²Ð½Ð¾Ð³Ð¾ ÑÑÑÐµÑÑÐ° Ð² ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ Ð¿ÑÐ¾Ð»Ð¸ÑÐµÑÐ°ÑÐ¸Ð¸ ÑÐ¼Ð±ÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½ÑÑ ÑÑÐ²Ð¾Ð»Ð¾Ð²ÑÑ ÐºÐ»ÐµÑÐ¾Ðº. Ð¦ÐµÐ»ÑÑ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð¾ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÑ Ð²Ð»Ð¸ÑÐ½Ð¸Ðµ ÑÐºÐ·Ð¾Ð³ÐµÐ½Ð½ÑÑ Ð»ÐµÐ½ÑÐ¸Ð²Ð¸ÑÑÑÐ½ÑÑ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¹, ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ¸Ñ LbNOX Ð¸ mitoLbNOX, Ð½Ð° Ð¿ÑÐ¾Ð»Ð¸ÑÐµÑÐ°ÑÐ¸Ñ Ð­Ð¡Ð, Ð½Ð¾ÐºÐ°ÑÑÐ½ÑÑ Ð¿Ð¾ Ð³ÐµÐ½Ñ Ndufs1. ÐÐ»Ñ Ð´Ð¾ÑÑÐ¸Ð¶ÐµÐ½Ð¸Ñ Ð¿Ð¾ÑÑÐ°Ð²Ð»ÐµÐ½Ð½Ð¾Ð¹ ÑÐµÐ»Ð¸ Ð±ÑÐ»Ð¾ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð¾ Ð¿ÐµÑÐµÐºÐ»Ð¾Ð½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ ÐºÐÐÐ LbNOX Ð¸ mitoLbNOX Ð² Ð»ÐµÐ½ÑÐ¸Ð²Ð¸ÑÑÑÐ½ÑÐ¹ Ð²ÐµÐºÑÐ¾Ñ, Ð¿ÑÐ¾Ð²ÐµÑÐºÐ° Ð²Ð»Ð¸ÑÐ½Ð¸Ñ Ð²ÑÐµÐ¼ÐµÐ½Ð½Ð¾Ð¹ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ ÑÑÐ¸Ñ Ð³ÐµÐ½Ð¾Ð² Ð½Ð° Ð¿ÑÐ¾Ð»Ð¸ÑÐµÑÐ°ÑÐ¸Ñ Ð­Ð¡Ð Ð¼ÑÑÐ¸, Ð½Ð¾ÐºÐ°ÑÑÐ½ÑÑ Ð¿Ð¾ Ndufs1, ÑÐ±Ð¾ÑÐºÐ° Ð²Ð¸ÑÑÑÐ½ÑÑ ÑÐ°ÑÑÐ¸Ñ Ñ ÐºÐÐÐ GFP Ð¸ Ð½Ð°ÑÐ¸Ñ Ð³ÐµÐ½Ð¾Ð², ÑÐ°ÑÑÑÑ Ð¸ÑÑÐ¸ÑÑÐ°. ÐÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð½ÑÐµ Ð¼ÐµÑÐ¾Ð´Ñ: ÑÑÐ°Ð½ÑÑÐ¾ÑÐ¼Ð°ÑÐ¸Ñ ÐºÐ»ÐµÑÐ¾Ðº E.coli, ÑÑÐ°Ð½ÑÑÐµÐºÑÐ¸Ñ Ð¸ ÑÑÐ°Ð½ÑÐ´ÑÐºÑÐ¸Ñ ÑÑÐºÐ°ÑÐ¸Ð¾ÑÐ¸ÑÐµÑÐºÐ¸Ñ ÐºÐ»ÐµÑÐ¾Ðº, ÑÐ±Ð¾ÑÐºÐ°, Ð¿Ð°ÐºÐ¾Ð²ÐºÐ° Ð¸ Ð¿Ð¾Ð´ÑÑÑÑ ÑÐ¸ÑÑÐ° Ð»ÐµÐ½ÑÐ¸Ð²Ð¸ÑÑÑÐ¾Ð², Ð¸Ð¼Ð¼ÑÐ½Ð¾ÑÐ¸ÑÐ¾ÑÐ¸Ð¼Ð¸ÑÐµÑÐºÐ¾Ðµ Ð¾ÐºÑÐ°ÑÐ¸Ð²Ð°Ð½Ð¸Ðµ, Ð¿ÑÐ¾ÑÐ¾ÑÐ½Ð°Ñ ÑÐ¸ÑÐ¾Ð¼ÐµÑÑÐ¸Ñ, Ð´Ð¸ÑÐ¿ÐµÑÑÐ¸Ð¾Ð½Ð½ÑÐ¹ Ð°Ð½Ð°Ð»Ð¸Ð· , T-ÐºÑÐ¸ÑÐµÑÐ¸Ð¹ Ð¡ÑÑÑÐ´ÐµÐ½ÑÐ°. Ð ÑÐ¾Ð´Ðµ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð¸ Ð¿Ð¾Ð»ÑÑÐµÐ½Ñ Ð»ÐµÐ½ÑÐ¸Ð²Ð¸ÑÑÑÐ½Ð¾ÑÐµ Ð²ÐµÐºÑÐ¾ÑÐ° Ñ ÐºÐÐÐ LbNOX Ð¸ mitoLbNOX, Ð±ÑÐ»Ð¾ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¾ Ð²Ð»Ð¸ÑÐ½Ð¸Ðµ Ð²ÑÐµÐ¼ÐµÐ½Ð½Ð¾Ð¹ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ Ð´Ð°Ð½Ð½ÑÑ Ð³ÐµÐ½Ð¾Ð² Ð² ÐºÐ¾Ð½ÑÑÐ¾Ð»ÑÐ½ÑÑ Ð¸ Ndufs1-/- Ð­Ð¡Ð Ð¼ÑÑÐ¸, Ð±ÑÐ» Ð¸Ð·Ð¼ÐµÑÐµÐ½ ÑÐ¸ÑÑ GFP ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐµÐ³Ð¾ Ð²Ð¸ÑÑÑÐ°. Ð ÐµÐ·ÑÐ»ÑÑÐ°ÑÑ Ð´Ð°Ð½Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÑ Ð¼Ð¾Ð³ÑÑ Ð±ÑÑÑ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ñ Ð´Ð»Ñ Ð´Ð°Ð»ÑÐ½ÐµÐ¹ÑÐ¸Ñ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ð¹ Ð² Ð¾Ð±Ð»Ð°ÑÑÐ¸ Ð¼ÐµÑÐ°Ð±Ð¾Ð»Ð¸Ð·Ð¼Ð° Ð­Ð¡Ð, ÐºÐ¾ÑÐ¾ÑÑÐµ ÑÐ»ÑÑÑÐ°ÑÑ Ð¿Ð¾Ð½Ð¸Ð¼Ð°Ð½Ð¸Ðµ Ð¼ÐµÑÐ°Ð½Ð¸Ð·Ð¼Ð¾Ð² ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ ÑÑÐ²Ð¾Ð»Ð¾Ð²ÑÑ ÐºÐ»ÐµÑÐ¾Ðº, ÑÑÐ¾ Ð²Ð°Ð¶Ð½Ð¾ Ð´Ð»Ñ ÑÐ°Ð·Ð²Ð¸ÑÐ¸Ñ ÐºÐ»ÐµÑÐ¾ÑÐ½ÑÑ ÑÐµÑÐ½Ð¾Ð»Ð¾Ð³Ð¸Ð¹ Ð¸ Ð¿ÑÐ°ÐºÑÐ¸ÑÐµÑÐºÐ¾Ð³Ð¾ Ð¿ÑÐ¸Ð¼ÐµÐ½ÐµÐ½Ð¸Ñ ÑÑÐ²Ð¾Ð»Ð¾Ð²ÑÑ ÐºÐ»ÐµÑÐ¾Ðº.
description: The given work is devoted to investigation of the role of reductive stress in the regulation of embryonic stem cell proliferation The goal of this work was to investigate the influence of exogenous lentivirus constructs that contain water-forming NADH oxidases LbNOX and mitoLbNOX on proliferative activity of mouse stem cells with Ndufs1-/-. To achieve the goal cloning LbNOX and mitoLbNOX cDNAs into lentivirus vector, check of temporary expression of LbNOX and mitoLbNOX in NDUFS1 gene knockout embryonic stem cells influence, lentivirus particle containing control GFP cDNA, LbNOX and mitoLbNOX cDNAs assembly and titer measurement were carried out. Methods that were used in this work: E.coli cells transformation , eukaryotic cell transfection and transduction, lentivirus particle assembly, titer measurement on HEK 293T cells and mESC, eukaryotic cell immunocytochemistry, flow cytometry, statistical analysis (ANOVA, Students t-test ). As a result of this work lentivirus based genetic constructs that contain LbNOX and mitoLbNOX cDNA were acquired, investigation of temporary expression of these genes in control and Ndufs1-/- mouse embryonic stem cells influence was performed, titer of control GFP containing virus was measured. Results of this work can be used as a base for further investigation in embryonic stem cell metabolism field. Research in this sphere improve our understanding of the stem cells functioning mechanisms which is important for introduction of this technology into the practice of regenerative medicine.