EGF and LIF enhanced the spontaneous reprogramming of Spermatogonial Stem Cells into Pluripotent State
Around 1x105 SSCs (cultured in medium 1 for 7 passages), Intermediate state cells (SSCs cultured in medium 1 for 5 passages and further cultured in medium 2 for 2 passages), GSPCs (SSCs cultured in medium 1 for 5 passages and further cultured in medium 2 for 5 passages) and ES-like cells (10 passages) were collected using the identical protocols. Total RNA was extracted using Trizol (Ambion Life Technologies) according to the Ambion standard RNA isolation procedure. mRNA was purified using the NEBNext Poly (A) mRNA Magnetic Isolation Beads (NEB, USA). Then the mRNA library was constructed with a NEBNext Ultra Directional RNA Library Prep Kit for Illumina (E7420S/L, NEB).