Unknown,Transcriptomics,Genomics,Proteomics

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Expression data of MACS sorted cells (SSEA-1+ and SSEA-1-) from porcine fetal fibroblast cell line

ABSTRACT: A subset of cells in cellular populations may be preferentially reprogrammed into pluripotent stem cells due to their innate gene expression characteristics. SSEA-1+ sorted cells were shown to be more amenable to reprogramming than other cells in porcine fetal fibroblast cell lines. This microarray data reveals which genes are unique to the SSEA1+ sorted cells and helps to give them an identity. Porcine fetal fibroblasts were subjected to Magnetic Activated Cell Sorting using anti-SSEA-1. Five biological replicates were performed resulting in 5 SSEA-1 positive fractions and 5 SSEA-1 negative fractions which were analyzed.

ORGANISM(S): Sus scrofa

SUBMITTER: Vanessa Hall

PROVIDER: E-GEOD-72025 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project 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.