GENERATION OF RAT EMBRYONIC GERM CELLS VIA INHIBITION OF TGFβ AND MEK PATHWAYS

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MOHAMMADI ALIREZA; ATTARI FARNOOSH; BABAPOUR VAHAB; HASSANI SEYEDEH NAFISEH; MASOUDI NAJMEHSADAT; SHAHVERDI ABDOLHOSSEIN; BAHARVAND HOSSEIN

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Journal: Cell Journal (Yakhteh) Year:2015 | Volume:17 | Issue:2 Page(s): 288-295

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Title

GENERATION OF RAT EMBRYONIC GERM CELLS VIA INHIBITION OF TGFβ AND MEK PATHWAYS

Author(s)

Keywords

PLURIPOTENCY

RAT

TGFβ PATHWAY

Abstract

Objective: Embryonic germ (EG) cells are the results of reprogramming primordial germ cells (PGC) in vitro. Studying these cells can be of benefit in determining the mechanism by which specialized cells acquire PLURIPOTENCY. Therefore in the current study we have tried to derive RAT EG cells with inhibition of transforming growth factor-b (TGFb) and mitogen-activated protein kinase kinase (MEK) signaling pathways.Materials and Methods: In this experimental study, RAT PGCs were cultured under feeder free condition with two small molecules that inhibited the above mentioned pathways. Under this condition only two-day presence of stem cell factor (SCF) as a survival factor was applied for PGC reprogramming. PLURIPOTENCY of the resultant EG cells were further confirmed by immunofluorescent staining, directed differentiation ability to neural and cardiac cells, and their contribution to teratoma formation as well. Moreover, chromosomal stability of two different EG cells were assessed through G-banding technique.Results: Formerly, derivation of RAT EG cells were observed solely in the presence of glycogen synthase kinase-3 (GSK3b) and MEK pathway inhibitors. Due to some drawbacks of inhibiting GSK3b molecules such as increases in chromosomal aberrations, in the present study we have attempted to assess a feeder-free protocol that derives EG cells by the simultaneous suppression of TGFb signaling and the MEK pathway. We have shown that RAT EG cells could be generated in the presence of two inhibitors that suppressed the above mentioned pathways. Of note, inhibition of TGFb instead of GSK3b significantly maintained chromosomal integrity. The resultant EG cells demonstrated the hallmarks of PLURIPOTENCY in protein expression level and also showed in vivo and in vitro differentiation capacities.Conclusion: RAT EG cells with higher karyotype stability establish from PGCs by inhibiting TGFb and MEK signaling pathways.

Cites

TEMPORAL GENE EXPRESSION AND DNA METHYLATION DURING EMBRYONIC STEM CELL DERIVATION

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APA: Copy

MOHAMMADI, ALIREZA, ATTARI, FARNOOSH, BABAPOUR, VAHAB, HASSANI, SEYEDEH NAFISEH, MASOUDI, NAJMEHSADAT, SHAHVERDI, ABDOLHOSSEIN, & BAHARVAND, HOSSEIN. (2015). GENERATION OF RAT EMBRYONIC GERM CELLS VIA INHIBITION OF TGFβ AND MEK PATHWAYS. CELL JOURNAL (YAKHTEH), 17(2), 288-295. SID. https://sid.ir/paper/644123/en

Vancouver: Copy

MOHAMMADI ALIREZA, ATTARI FARNOOSH, BABAPOUR VAHAB, HASSANI SEYEDEH NAFISEH, MASOUDI NAJMEHSADAT, SHAHVERDI ABDOLHOSSEIN, BAHARVAND HOSSEIN. GENERATION OF RAT EMBRYONIC GERM CELLS VIA INHIBITION OF TGFβ AND MEK PATHWAYS. CELL JOURNAL (YAKHTEH)[Internet]. 2015;17(2):288-295. Available from: https://sid.ir/paper/644123/en

IEEE: Copy

ALIREZA MOHAMMADI, FARNOOSH ATTARI, VAHAB BABAPOUR, SEYEDEH NAFISEH HASSANI, NAJMEHSADAT MASOUDI, ABDOLHOSSEIN SHAHVERDI, and HOSSEIN BAHARVAND, “GENERATION OF RAT EMBRYONIC GERM CELLS VIA INHIBITION OF TGFβ AND MEK PATHWAYS,” CELL JOURNAL (YAKHTEH), vol. 17, no. 2, pp. 288–295, 2015, [Online]. Available: https://sid.ir/paper/644123/en

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