|Application:||Gene expression microarray analysis|
|Number of samples:||5|
|Release date:||Jul 10 2009|
|Last update date:||May 3 2013|
|Dataset link||Induced Pluripotent Stem Cells from the Pig Somatic Cells|
The open reading frames of the human (h) SOX2, hKLF4, and hc-MYC have been cloned into the BamHI and EcoRI sites of the lentiviral vector, FUGW and the hOCT4 cDNA into the pSIN18.cPPT.hEF1a.EGFP.WPRE vector at its BamHI and SalI sites. Pseudovirus was produced in human 293FT cells (Invitrogen) by transfection with each lentiviral vector (FUGW or pSIN18) along with VSV-G envelope vector (pMD2.G) and packaging vector (psPAX2) by using Lipofectamine-Plus reagents (Invitrogen). The target cells were porcine fibroblasts expressing enhanced green fluorescent protein (EGFP-PFF) derived at day 34 of pregnancy. Titered virus was used to infect the target cells (1 x 10^5 /35mm dish). On the second day following infection (day 2), the cells were dispersed with trypsin and transferred to 10 cm plates preseeded with irradiated mouse embryonic fibroblasts (MEF), and after day 3 the cells were maintained on a culture medium standardized for human ESC containing 4 ng/ml human FGF2.