Analysis of cell growth and gene expression of porcine adipose tissue‐derived mesenchymal stem cells as nuclear donor cell

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.

Porcine somatic cell nuclear transfer donor sources affect transplant success: A meta-analysis

“…Two studies included donor adults that were 4 years-of-age and one study used the Meishan pig species. Five studies included NCSU-23 (Wang et al 2015;Hua et al 2016) and others used Porcine zygote medium (PZM) (Liu et al 2012;Oh et al 2014;Liu et al 2015).…”

“…Existing reports suggest that porcine fetal fibroblasts can be used as donor cells for SCNT and donor nuclei can be taken from embryonic stem and germ cells, induced pluripotent stem cells, adult stem cells and fetal and neonatal or adult somatic cells. However, whether differentiation of early cell lines offers better cloning efficiency is not clear (Gade et al 2012;Liu et al 2012;Oh et al 2014). At present, most studies use pig fetal fibroblasts as donor cells Hua et al 2016).…”

“…Defects of somatic cell nuclear epigenetic reprogramming cause SCNT embryo death (Salilew-Wondim et al 2013), so removal of DNA methylation and histone acetylation may increase SCNT blastocyst formation (Lan et al 2017). It has been found that adult donors of 4 yearsof-age are optimal (Liu et al 2012;Oh et al 2014) and SCNT blastocyst formation between adult and fetal donors differ.…”

“…Using SCNT cloning, porcine herd boars should increase, and this is of economic value. Because cloning herd boars requires adult donor cells, and research is conflicting about whether fetal cells are viable sources for SCNT (Liu et al 2012;Oh et al 2014;Liu et al 2015;Hua et al 2016), we analyzed the influence of donor cells on pig SCNT by specifically comparing fetal and adult donor sources. Publications meeting inclusion and exclusion criteria are depicted in Fig 1 and Table 1.…”

Enhanced insulin production and reprogramming efficiency of mesenchymal stem cells derived from porcine pancreas using suitable induction medium

“…[9][10][11][12] pMSCs have been reported to possess a broad differentiation ability, and in suitable induction media, they can differentiate not only into similar lineage cell types, including adipocytes, osteocytes, and chondrocytes, 8 but also into different lineage cell types, such as endothelial cells, 13,14 hepatocytes, 14 and neural cells. 15,16 Interestingly, although MSCs have been characterized by differentiation into various cell types, the cells differ in their proliferative capacity, 17 pluripotency marker profile, 17 morphology, 9 immunophenotype, 9 and differentiation potential depending on their tissue of origin. 9,18 In the case of pancreas-derived MSCs, islet-or exocrine cell-specific genes are expressed.…”