Reprogramming of ovine adult fibroblasts to pluripotency via drug-inducible expression of defined factors.
Summary of "Reprogramming of ovine adult fibroblasts to pluripotency via drug-inducible expression of defined factors."
Reprogramming of somatic cells in the enucleated egg made Dolly, the sheep, the first successfully cloned mammal in 1996. However, the mechanism of sheep somatic cell reprogramming has not yet been addressed. Moreover, sheep embryonic stem (ES) cells are still not available, which limits the generation of precise gene-modified sheep. In this study, we report that sheep somatic cells can be directly reprogrammed to induced pluripotent stem (iPS) cells using defined factors (Oct4, Sox2, c-Myc, Klf4, Nanog, Lin28, SV40 large T and hTERT). Our observations indicated that somatic cells from sheep are more difficult to reprogram than somatic cells from other species, in which iPS cells have been reported. We demonstrated that sheep iPS cells express ES cell markers, including alkaline phosphatase, Oct4, Nanog, Sox2, Rex1, stage-specific embryonic antigen-1, TRA-1-60, TRA-1-81 and E-cadherin. Sheep iPS cells exhibited normal karyotypes and were able to differentiate into all three germ layers both in vitro and in teratomas. Our study may help to reveal the mechanism of somatic cell reprogramming in sheep and provide a platform to explore the culture conditions for sheep ES cells. Moreover, sheep iPS cells may be directly used to generate precise gene-modified sheep.Cell Research advance online publication 11 January 2011; doi:10.1038/cr.2011.6.
Affiliation
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Journal Details
This article was published in the following journal.
Name: Cell research
ISSN: 1748-7838
Pages:
Links
- PubMed Source: http://www.ncbi.nlm.nih.gov/pubmed/21221129
- DOI: http://dx.doi.org/10.1038/cr.2011.6
Medical and Biotech [MESH] Definitions
Nuclear Reprogramming
The process that reverts CELL NUCLEI of fully differentiated somatic cells to a pluripotent or totipotent state. This process can be achieved to a certain extent by NUCLEAR TRANSFER TECHNIQUES, such as fusing somatic cell nuclei with enucleated pluripotent embryonic stem cells or enucleated totipotent oocytes. GENE EXPRESSION PROFILING of the fused hybrid cells is used to determine the degree of reprogramming. Dramatic results of nuclear reprogramming include the generation of cloned mammals, such as Dolly the sheep in 1997.
Visna-maedi Virus
A species of LENTIVIRUS, subgenus ovine-caprine lentiviruses (LENTIVIRUSES, OVINE-CAPRINE), that can cause chronic pneumonia (maedi), mastitis, arthritis, and encephalomyelitis (visna) in sheep. Maedi is a progressive pneumonia of sheep which is similar to but not the same as jaagsiekte (PULMONARY ADENOMATOSIS, OVINE). Visna is a demyelinating leukoencephalomyelitis of sheep which is similar to but not the same as SCRAPIE.
Arthritis-encephalitis Virus, Caprine
A species of LENTIVIRUS, subgenus ovine-caprine lentiviruses (LENTIVIRUSES, OVINE-CAPRINE), closely related to VISNA-MAEDI VIRUS and causing acute encephalomyelitis; chronic arthritis; PNEUMONIA; MASTITIS; and GLOMERULONEPHRITIS in goats. It is transmitted mainly in the colostrum and milk.
Nitric Oxide Synthase Type Ii
A CALCIUM-independent subtype of nitric oxide synthase that may play a role in immune function. It is an inducible enzyme whose expression is transcriptionally regulated by a variety of CYTOKINES.
Hypoxia-inducible Factor 1
A basic helix-loop-helix transcription factor that plays a role in APOPTOSIS. It is composed of two subunits: ARYL HYDROCARBON RECEPTOR NUCLEAR TRANSLOCATOR and HYPOXIA-INDUCIBLE FACTOR 1, ALPHA SUBUNIT.
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