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A novel feeder-free system for human embryonic stem cells and characterization of their sublines with autogenic and allogenic cultivationстатья
Информация о цитировании статьи получена из
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 21 августа 2017 г.
- Авторы: Koltsova A.M., Voronkina I.V., Gordeeva O.F., Zenin V.V., Lifantseva N.V., Musorina A.S., Smagina L.V., Jakovleva T.K., Poljanskaya G.G.
- Журнал: Cell and Tissue Biology
- Том: 7
- Номер: 1
- Год издания: 2013
- Издательство: Maik Nauka/Interperiodica Publishing
- Местоположение издательства: Russian Federation
- Первая страница: 637
- Последняя страница: 652
- DOI: 10.1134/S1990519X13010094
- Аннотация: We developed a feederfree system for human embryonic stem cells (ESCs) based on extracellular matrix protein (ECM) as the substrate. ECM was synthesized by mesenchymal stem cells (SC5MSC) derived from an original ESC line, SC5. The ECM proteins fibronectin and laminin facilitate ESC growth in the feederfree system. An important component of this system is a conditioned medium from SC5MSC cells. Two ESC sublines were obtained: SC5FF cells were cultured in an autogenic, and SC7FF in an allo genic, feederfree system. SC5FF and SC7FF underwent more than 300 and 115 population doublings, respectively, and retain a normal diploid karyotype. Histochemical and immunofluorescence assays showed that both sublines express undifferentiated ESC markers—alkaline phosphatase, Oct4, SSEA4, and TRA181—as well as multidrug resistance transporter ABCG2. PCR assay revealed that undifferenti ated SC5FF cells, like the original SC5 line, maintained on feeder cells express OCT4 and NANOG genes common for somatic cells and DPPA3/STELLA and DAZL genes common for germ line cells. Expression of these genes was gradually diminished during differentiation of embryoid bodies, whereas expression of genes specific for early differentiated cells increased: GATA4, AFP (extraembryonic and embryonic endoderm), PAX6 (neuroectoderm), and BRY (mesoderm). ESC properties (karyotype structure, average time of popu lation doubling, undifferentiated cell number in population) of the SC5 and SC7 and SC5FF and SC7FF sublines derived from original ESCs were not altered. It shows that the feederfree systems, which are more stable than any feeder systems, maintain key ESC properties and may be recommended for fundamental, bio medical, and pharmacological studies performed with human ESCs.
- Добавил в систему: Воронкина Ирина Владимировна