Piglets cloned from induced pluripotent stem cells

Abstract

Figure 1Derivation and characterization of cloned piglets from piPSCs. (A) Preimplantation and post-implantation development of the cloned embryos from piPSCs. Embryos at two-cell (a), four-cell (b), eight-cell (c), blastocyst stages (d, e) and two 36 day-old cloned fetuses (f) are shown. Scale bars are 100 μm. (B) The morphology and fluorescence of the hooves (left), tails (middle) and fibroblasts (right) of the 36 day-old embryos. Scale bars are 100 μm. (C) The morphology, fluorescence and hematoxylin/eosin-stained sections of tissues from piglet 00536-3#. Scale bars are 100 μm. (D) Cloned piglets. 00507-4# from differentiated iPF4-2 cell, 4 days old; 227-1#, 2#, 3# from undifferentiated iPF4-2, 2 days old. (E) Porcine ear fibroblasts (PEFs) from 00507-4#, EGFP positive. Scale bars are 100 μm. (F) PCR demonstrating genomic integration of Oct4, Sox2, and EGFP using tissues of the cloned fetuses and piglets. PEF, the original fibroblasts used to create iPF4-2. 00518, 00536, 00507, 227, foster mothers. 00518-1#, 00518-2#, the cloned fetuses derived from differentiated iPF4-2 cells. 00536-3# , 00507-4#, the cloned piglets derived from iPF4-2-differentiated cells. 227-1#~4#, the HMC piglets derived from iPF4-2. (G) Microsatellite analysis of the donor piPSC line iPF4-2, cloned fetuses and piglets. 00518, 00536, 00507, 227, foster mothers; 00518-1# and 00518-2#, cloned fetuses; 00536-3# and 00507-4#, the cloned piglets from differentiated iPF4-2 cells; 227-1#~4#, the cloned piglets derived from the Scriptaid-treated NT embryos from iPF4-2 cells.www.cell-research.com | Cell Research Nana Fan et al. 165npg

Germline transgenesis in pigs by cytoplasmic microinjection of Sleeping Beauty transposons

“…Recently, several groups described the derivation of porcine induced pluripotent cells (iPS) [16][17][18][19][20] , however, only low chimerism has been found after blastocyst complementation experiments 21,22 . The employment of porcine iPS cells in the SCNT method resulted in extremely low rates of born piglets 23 .…”

Factors Determining the Efficiency of Porcine Somatic Cell Nuclear Transfer: Data Analysis with Over 200,000 Reconstructed Embryos

“…In recent studies, to correct or relieve the incomplete epigenetic reprogramming of cloned embryos, different cell types were used as the nuclear donor for pig SCNT, such as fetal fibroblasts (FFs; Onishi et al, 2000), preadipocytes (Tomii et al, 2005), adult mesenchymal stem cells (MSCs; Faast et al, 2006), recloned pig somatic cells (Cho et al, 2007), and induced pluripotent stem cells (iPSCs; Fan et al, 2013).…”

Characterization of porcine partially reprogrammed iPSCs from adipose-derived stem cells

“…High proliferation activity of pPiPSCs and their differentiated cells may satisfy the long-term requirement of in vitro cellular drug screening, although further studies are required to confirm this. During SCNT from iPSCs, insufficient silencing of exogenous genes would compromise the development of reconstructive embryos (Fan et al 2013). Thus, differentiated cells from pPiPSCs and pFiPSCs were used for SCNT.…”

“…West et al (2011) established chimeric pigs from mesenchymal stem cell (MSC)-derived iPSCs, and, although this was controversial, it has been suggested that MSC-derived iPSCs may be beneficial for the generation of porcine offspring. Fan et al (2013) studied iPSCs and differentiated porcine iPSCs, and concluded that the expressions of exogenous transcription factors in porcine iPSCs have a negative effect on the production of iPSC-derived clone pigs. Accordingly, we wonder whether pADSC-derived PiPSCs with the perfect regulation exogenous gene expression are beneficial for the development of SCNT embryos.…”