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E-GEOD-15472 - Transcription profiling of pig induced pluripotent stem cells from somatic cells

Status
Submitted on 31 March 2009, released on 19 July 2009, last updated on 1 May 2014
Organism
Sus scrofa
Samples (5)
Array (1)
Protocols (6)
Description
We have derived induced porcine pluripotent stem cells (iPPSCs) from porcine fetal fibroblasts by lentiviral transduction of four human (h) reprogramming genes, hOCT4, hSOX2, hKLF4 and hc-MYC , the same combination of factors used for deriving induced pluripotent stem cell (iPSC) lines in both mouse and human. The obtained iPPSC lines resemble human embryonic stem cells (ESC) in their gross morphology and dependence on FGF2, on the other hand, the iPPSCs share characteristics like growth rate and cell surface markers with mESC . Additionally, the iPPSCs express pluripotency- associated genes similar to mouse and human iPSCs as well as ESC, along with the pig epiblast cells. Some of the iPPSC lines retained a stable karyotype and phenotype even in culture for a prolonged period of time (passage 39). The iPPSCs can be induced to differentiate along lineages representative of the three embryonic germ layers both in vitro and in vivo demonstrating the pluripotency of these cells. Experiment Overall Design: 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.
Experiment types
transcription profiling by array, unknown experiment type
Contact
Citation
Derivation of induced pluripotent stem cells from pig somatic cells. Toshihiko Ezashi, Bhanu Prakash V L Telugu, Andrei P Alexenko, Shrikesh Sachdev, Sunilima Sinha, R Michael Roberts.
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-15472.idf.txt
Sample and data relationshipE-GEOD-15472.sdrf.txt
Raw data (1)E-GEOD-15472.raw.1.zip
Processed data (1)E-GEOD-15472.processed.1.zip
Array designA-AFFY-75.adf.txt
R ExpressionSetE-GEOD-15472.eSet.r
Links