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Murine Fibroblast Cell Lines, NIH-3T3 and STO,Cannot be Reprogrammed to Pluripotent Stateстатья
Статья опубликована в журнале из списка RSCI Web of Science
Статья опубликована в журнале из перечня ВАКСтатья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 11 апреля 2019 г.
- Авторы: Skvortsova E.V., Sinenko S.A., Tomilin A.N.
- Журнал: Онтогенез
- Том: 49
- Номер: 4
- Год издания: 2018
- Первая страница: 45
- Последняя страница: 45
- DOI: 10.1134/S0475145018010081
- Аннотация: To date different cell types of various mammalian species have been reprogrammed by Yamanaka’s cocktail of transcription factors, includ- ing OCT4, KLF4, SOX2, and C-MYC. It has been al- so shown that several primary human cancer cell lines were reprogrammed to induced pluripotent stem cells (iPSCs). We sought if widely used mouse fibroblast cell lines could be reprogrammed to iPSCs. This ap- proach of generating iPSCs from stable murine cell lines assumed to be valuable for different experimental settings as it allows generating CRISPER/CAS9 base mutant cell lines that can be assayed in reprogram- ming. At the other hand only single study has reported reprogramming murine carcinoma cell line. To this regard we investigated reprogramming two fibroblast- originated cell lines, NIH-3T3 and STO, with use of highly effective lentiviral polycistronic OKSM expres- sion system. The reprogramming of wild type murine embryonic fibroblasts and NIH-3T3 and STO cells were performed with the conventional method with use of feeder embryonic fibroblasts and N2B27 2i me- dia. Two independent reprogramming experiments have shown that in contrast to control wild-type fibro- blasts, NIH-3T3 and STO cells could not be repro- grammed to pluripotent state. While in all groups has been seen generation of cell clones, NIH-3T3 and STO clones were developed much later then wild type ones, and they contained round and larger cells, which are not typical for iPSCs colonies. Alkaline phospha- tase and immunostaining on Nanog and SSEA1 con- firmed that the cell colonies derived from NIH-3T3 and STO did not express these pluripotency markers. Furthermore in contrast to wild-type fibroblast de- rived iPSC clones, NIH-3T3 and STO clones could not be maintained by multiple cell passaging in em- bryonic cell media. Thus this data suggest that proba- bly genetic abnormalities or abnormal cell signaling or both could make these immortalized cell lines be re- fractory to reprogramming to pluripotent state.
- Добавил в систему: Скворцова Елена Вячеславовна