Porcine embryonic stem cells: Facts, challenges and hopes

Brevini, Tiziana A. L.; Abednatanzi, Sara; Cillo, F.; Crestan, M.; Gandolfi, F.

doi:10.1016/j.theriogenology.2007.05.043

Cited by 100 publications

(82 citation statements)

References 55 publications

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“…ESCs have been considered as a useful tool for generating transgenic pig. However, the research of these potential applications progressed slowly, because no authentic porcine ESCs is available to date (Li et al, 2004;Brevini et al, 2007;Kim et al, 2007;Hall, 2008;Kim et al, 2010). Although three groups announced recently they had established iPSCs in pig by introduced human or mouse derived transcriptional factors (Oct4, Sox2, Klf4, and C-myc), none of them successfully applied the iPSCs in generating chimeras, which is the basic standard to define pluripotency (Esteban et al, 2009;Ezashi et al, 2009;Wu et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Overexpression Nanog Activates Pluripotent Genes in Porcine Fetal Fibroblasts and Nuclear Transfer Embryos

Zhang

Luo

Bou

et al. 2011

The Anatomical Record

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Nanog as an important transcription factor plays a pivotal role in maintaining pluripotency and in reprogramming the epigenome of somatic cells. Its ability to function on committed somatic cells and embryos has been well defined in mouse and human, but rarely in pig. To better understand Nanog's function on reprogramming in porcine fetal fibroblast (PFF) and nuclear transfer (NT) embryo, we cloned porcine Nanog CDS and constructed pcDNA3.1 (þ)/Nanog and pEGFP-C1/Nanog overexpression vectors and transfected them into PFFs. We studied the cell biological changes and the expression of Nanog, Oct4, Sox2, Klf4, C-myc, and Sall4 in transfected PFFs. We also detected the development potential of the cloned embryos harboring Nanog stably overexpressed fibroblasts and the expression of Oct4, Sox2, and both endogenous and exogenous Nanog in these embryos. The results showed that transient overexpression Nanog in PFF could activate the expression of Oct4 (5-fold), C-myc (2-fold), and Sall4 (5-fold) in somatic cells, but they could not be maintained during G418 selection. In NT embryos, although Nanog overexpression did not have a significant effect on blastocyst development rate and blastocyst cell number, it could significantly activate the expression of endogenous Nanog, Oct4, Sox2 to 160-fold, 93-fold, and 182-fold, respectively (P < 0.05). Our results demonstrate that Nanog could interact with and activate other pluripotent genes both in PFFs and embryos. Anat Rec, 294:1809Rec, 294: -1817Rec, 294: , 2011. V V C 2011 Wiley-Liss, Inc.

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Section: Introductionmentioning

confidence: 99%

Overexpression Nanog Activates Pluripotent Genes in Porcine Fetal Fibroblasts and Nuclear Transfer Embryos

Zhang

Luo

Bou

et al. 2011

The Anatomical Record

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“…This discovery also heralded the prospects of using ESC in regenerative medicine. Despite their undoubted promise as sources of tissue transplants, many road blocks remain to using human ESC as a source of transplant material, especially as a means to test the efficacy of therapies and the safety of the transferred cells in animals whose anatomy and physiology better resemble the human than the mouse (11)(12)(13)(14)(15). The pig is a potentially useful model in this regard because of similarities in organ size, immunology, and whole animal physiology (16)(17)(18).…”

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confidence: 99%

“…There has been a similar lack of success with other ungulate species. The earliest reports announcing the derivation of ESC-like cells from ICM of pigs appeared in the early 1990s (19)(20)(21), but these ESC-like cells and many others since then, including ones for cattle, goat, and sheep, as well as for pig, have failed to meet the full criteria to define them as ESC (11)(12)(13). There are a number of reasons that might explain the problems encountered, including choice of the wrong stage of embryo development to establish the cultures, inappropriate culture and cell passage conditions, and contamination by more vigorously growing cells, such as the endoderm and trophectoderm of the blastocyst from which the culture was derived.…”

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confidence: 99%

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Derivation of induced pluripotent stem cells from pig somatic cells

Ezashi¹,

Telugu²,

Alexenko³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

439

399

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For reasons that are unclear the production of embryonic stem cells from ungulates has proved elusive. Here, we describe induced pluripotent stem cells (iPSC) derived from porcine fetal fibroblasts by lentiviral transduction of 4 human (h) genes, hOCT4, hSOX2, hKLF4, and hc-MYC, the combination commonly used to create iPSC in mouse and human. Cells were cultured on irradiated mouse embryonic fibroblasts (MEF) and in medium supplemented with knockout serum replacement and FGF2. Compact colonies of alkaline phosphatase-positive cells emerged after Ϸ22 days, providing an overall reprogramming efficiency of Ϸ0.1%. The cells expressed porcine OCT4, NANOG, and SOX2 and had high telomerase activity, but also continued to express the 4 human transgenes. Unlike human ESC, the porcine iPSC (piPSC) were positive for SSEA-1, but negative for SSEA-3 and -4. Transcriptional profiling on Affymetrix (porcine) microarrays and real time RT-PCR supported the conclusion that reprogramming to pluripotency was complete. One cell line, ID6, had a normal karyotype, a cell doubling time of Ϸ17 h, and has been maintained through >220 doublings. The ID6 line formed embryoid bodies, expressing genes representing all 3 germ layers when cultured under differentiating conditions, and teratomas containing tissues of ectoderm, mesoderm, and endoderm origin in nude mice. We conclude that porcine somatic cells can be reprogrammed to form piPSC. Such cell lines derived from individual animals could provide a means for testing the safety and efficacy of stem cell-derived tissue grafts when returned to the same pigs at a later age.iPS ͉ reprogramming ͉ OCT4

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“…Recently feeder-free system has been adapted especially for culturing hES cells to eliminate the potential issues of infection and cross-species contamination [6,7]. Some feeder-free systems support the culture of undifferentiated hES cells more efficiently than others [8,9], however, porcine and bovine epiblast and ICM cells cultured on a feeder free system did not grow continuously and started senescence or differentiation [10][11][12][13]. In case of the farm animals, ES cell-like cells are also commonly cultured on feeder cells because the molecular pathways and key molecules required in maintaining pluripotency in these species are still unknown [14].…”

Section: Introductionmentioning

confidence: 99%

Derivation of buffalo embryonic stem-like cells from in vitro-produced blastocysts on homologous and heterologous feeder cells

Kumar

Anand

Singh

et al. 2011

J Assist Reprod Genet

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Purpose The aim of the present study is to compare the ability of homologous and heterologous embryonic fibroblast feeder layers to support isolation and proliferation of buffalo ES-like cells generated from hatched and expanded blastocysts produced by in vitro fertilization and characterization of derived cells through expression of pluripotent markers. Methods Embryonic stem cells were derived from hatched and expanded blastocysts through intact blastocyst culture and enzymatic method respectively and compared for proliferation rate on homologous (buffalo) and heterologous feeder layers (goat and sheep). Results A total of 69 hatched and 83 expanded blastocysts were used for isolation of inner cell masses which were seeded on buffalo, goat and sheep embryonic feeder layers. Following seeding, attachment rate, primary colony formation rate and survival to maximum number of passages were observed to be higher on homologous feeder layers.Conclusions Upon comparison of different feeder layer cells for derivation and maintenance of buffalo ES-like cells from hatched and expanded blastocysts, buffalo embryonic fibroblast cells were able to provide a better environment for maintaining pluripotency in culture conditions.

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Porcine embryonic stem cells: Facts, challenges and hopes

Cited by 100 publications

References 55 publications

Overexpression Nanog Activates Pluripotent Genes in Porcine Fetal Fibroblasts and Nuclear Transfer Embryos

Overexpression Nanog Activates Pluripotent Genes in Porcine Fetal Fibroblasts and Nuclear Transfer Embryos

Derivation of induced pluripotent stem cells from pig somatic cells

Derivation of buffalo embryonic stem-like cells from in vitro-produced blastocysts on homologous and heterologous feeder cells

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