scbmtБИОМЕДИЦИНАJournal Biomed2074-59822713-0428Scientific center of biomedical technologies of Federal Medical and Biological Agencyscbmt-65Research ArticleМЕТОДЫ БИОМЕДИЦИНСКИХ ИССЛЕДОВАНИЙMETHODS OF BIOMEDICAL RESEARCHESВыживаемость NGF-модифицированных эмбриональных стволовых клеток мыши в составе многоклеточной бластоцистыThe survival of NGF-modified embryonic stem cell in mouse multicellular blastocystМежевикинаЛ. М.MezhevikinaL. M.mezhevikina@rambler.ruХрамцоваЕ. А.KhramtsovaE. A.noemail@neicon.ruФесенкоЕ. Е.FesenkoE. E.noemail@neicon.ruФГБУН Институт биофизики клетки РАНRussian Federation201726022019023344Copyright © Межевикина Л.М., Храмцова Е.А., Фесенко Е.Е., 20192019Межевикина Л.М., Храмцова Е.А., Фесенко Е.Е.Mezhevikina L.M., Khramtsova E.A., Fesenko E.E.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.scbmt.ru/jour/article/view/65

Исследовано влияние микроокружения на рост и выживаемость NGF-модифицированных эмбриональных стволовых клеток (NGF-ЭСК) мыши с экспрессией зеленого флуоресцирующего белка GFP в составе многоклеточной бластоцисты. Показано, что после микроинъекции NGF-ЭСК мигрируют и встраиваются преимущественно в область расположения клеток внутренней клеточной массы (ВКМ). Они продуцируют NGF и GFP и могут пролиферировать в составе химерной бластоцисты. При увеличении длительности культивирования химерных бластоцист до 120 ч наблюдается снижение интенсивности флуоресценции белка GFP, секреция NGF не обнаруживается. Полагаем, что это происходит в результате перепрограммирования NGF-ЭСК под влиянием клеток ВКМ, в результате чего подавляется экспрессия NGF/GFP и, следовательно, преждевременная дифференцировка ЭСК в направлении нейроэктодермы. Полученные результаты свидетельствуют о том, что микроокружение и клеточные взаимодействия играют решающую роль в процессах перехода NGF-ЭСК из состояния плюрипотентности к нейральной дифференцировке, которая является базовой на ранних стадиях развития.

We investigated the effect of the microenvironment on the growth and survival of NGF-modified embryonic stem cells (NGF-ESC) with the expression of green fluorescent protein GFP in mouse multicellular blastocyst. Our results show that after the microinjection (MI), NGF-ESC migrate and co-culture primarily in the region of inner cell mass (ICM). They produce NGF and GFP and are capable for proliferation in chimeric blastocyst. However, if the cultivation period is prolonged to 120 hours, the decrease in the intensity of GFP fluorescence strength and NGF secretion is observed. We conjecture that this occurs as a result of reprogramming of NGF-ESC under the influence of ICM leading to the suppression of NGF/GFP expression and, therefore, earlier differentiation of ESC towards the neuroectoderm. Obtained results indicate that the microenvironment and cellular interaction play a key role in transferring NGF-ESC from the state of pluripotency to the neutral differentiation which is basic during the early stages of development.

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The authors declare that there are no conflicts of interest present.