Transcriptional profiling of LIF and FGF-2 signaling pathways in buffalo embryonic stem cells derived from in vitro fertilized, parthenogenetic and handmade cloned embryos

Document Type : Full Research Paper

Authors

Iranian Research Organization for Science and Technology (IROST).

Abstract

Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of blastocyst and differentiate into all three embryonic germ layers: ectoderm, endoderm, and mesoderm. There is less information available about the factors that are affecting buffalo ES cells in culture. In this study, expression profiles of the Leukemia Inhibitory Factor (LIF) and basic Fibroblast Growth Factor (FGF-2) signaling pathways were investigated to better understand the relationships of the signaling pathways for self-renewal in buffalo ES cells. Buffalo ES cells were derived from in vitro fertilized (iESC), parthenogenetic (pESC) and handmade cloned (cESC) embryos. Alkaline phosphatase and immune-fluorescence staining were used to characterize buffalo ES cells. Feeder layer was used for ESCs culture, and culture medium consisting of Knockout- Dulbecco’s Modified Eagle’s Medium (Ko-DMEM) supplemented with Knockout Serum Replacement (KSR), leukemia inhibitory factor (LIF), basic fibroblast growth factor-2 (FGF-2), L-glutamine, nonessential amino acids and gentamicin. Gene expression was analysed by RT-PCR for signaling pathways. Results showed that, the expression of FGF-2 was higher than LIF in buffalo ESCs. LIF, FGF, their receptors and intermediate signaling pathways was expressed at almost same level in three sources of buffalo ESCs.

Keywords

References

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History

  • Receive Date: 21 April 2013
  • Revise Date: 23 July 2013
  • Accept Date: 23 August 2014

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