Analysis of cell growth and gene expression of porcine adipose tissue‐derived mesenchymal stem cells as nuclear donor cell |
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Authors: | Hyun Ju Oh Jung Eun Park Eun Jung Park Min Jung Kim Geon A Kim Sang Ho Rhee Sang Hyun Lim Sung Keun Kang Byeong Chun Lee |
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Institution: | 1. Department of Theriogenology and Biotechnology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University;2. College of Liberal Arts and Sciences, University of Washington, , Seattle, Washington, USA;3. Central research institute, Stem Cell Research Institute, K‐Stem Cell, , Seoul, 153‐768 Korea |
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Abstract: | In several laboratory animals and humans, adipose tissue‐derived mesenchymal stem cells (ASC) are of considerable interest because they are easy to harvest and can generate a huge proliferation of cells from a small quantity of fat. In this study, we investigated: (i) the expression patterns of reprogramming‐related genes in porcine ASC; and (ii) whether ASC can be a suitable donor cell type for generating cloned pigs. For these experiments, ASC, adult skin fibroblasts (AF) and fetal fibroblasts (FF) were derived from a 4‐year‐old female miniature pig. The ASC expressed cell‐surface markers characteristic of stem cells, and underwent in vitro differentiation when exposed to specific differentiation‐inducing conditions. Expression of DNA methyltransferase (DNMT)1 in ASC was similar to that in AF, but the highest expression of the DNMT3B gene was observed in ASC. The expression of OCT4 was significantly higher in FF and ASC than in AF (P < 0.05), and SOX2 showed significantly higher expression in ASC than in the other two cell types (P < 0.05). After somatic cell nuclear transfer (SCNT), the development rate of cloned embryos derived from ASC was comparable to the development of those derived using FF. Total cell numbers of blastocysts derived using ASC and FF were significantly higher than in embryos made with AF. The results demonstrated that ASC used for SCNT have a potential comparable to those of AF and FF in terms of embryo in vitro development and blastocyst formation. |
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Keywords: | adipose tissue‐derived mesenchymal stem cells adult fibroblasts fetal fibroblasts gene expression porcine somatic cell nuclear transfer |
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