Putative embryonic stem cells derived from porcine cloned blastocysts using induced pluripotent stem cells as donors

Kim, Eunhye; Hwang, Seon‐Ung; Yoo, Hyunju; Yoon, Jong Hwan; Jeon, Yubyeol; Kim, Hyunggee; Jeung, Eui Bae; Lee, Chang Kyu; Hyun, Sang Hwan

doi:10.1016/j.theriogenology.2015.09.051

Cited by 20 publications

(23 citation statements)

References 80 publications

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“…All of the pESC lines were established and characterized in a previous study [ 30 ]. In brief, hatched porcine blastocysts were obtained by IVF and SCNT using in vitro matured (IVM) oocytes.…”

Section: Methodsmentioning

confidence: 99%

Ultrastructural comparison of porcine putative embryonic stem cells derived by <i>in vitro</i> fertilization and somatic cell nuclear transfer

Yoo

Kim

Hwang

et al. 2016

Journal of Reproduction and Development

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The ultrastructure of porcine putative embryonic stem cells and porcine fetal fibroblasts (PFFs) was analyzed by transmission electron microscopy. The aim of this study was to compare the features of organelles in in vitro fertilization (IVF) derived porcine embryonic stem cells (IVF-pESCs) and somatic cell nuclear transfer (SCNT) derived pESCs (SCNT-pESCs). Also, the features of organelles in high-passage IVF-pESCs were compared with those in low-passage cells. The ultrastructure of PFFs showed rare microvilli on the cell surfaces, polygonal or irregular nuclei with one to two reticular-shaped nucleoli and euchromatin, low cytoplasm-to-nucleus ratios, rare ribosomes, rare rough endoplasmic reticulum, elongated mitochondria, rich lysosomes and rich phagocytic vacuoles. IVF-pESCs showed rare microvilli on the cell surfaces, round or irregular nuclei with one to two reticular-shaped nucleoli and euchromatin, low cytoplasm-to-nucleus ratios, rich ribosomes, long stacks of rough endoplasmic reticulum, elongated mitochondria, rare lysosomes and rare autophagic vacuoles. By contrast, SCNT-pESCs showed rich microvilli with various lengths and frequencies on the cell surfaces, polygonal nuclei with one reticular shaped nucleoli and heterochromatin, high cytoplasm-to-nucleus ratios, rare ribosomes, rare rough endoplasmic reticulum, round mitochondria, rich lysosomes and rich phagocytic vacuoles with clear intercellular junctions. Furthermore, high-passage IVF-pESCs showed irregularly shaped colonies, pyknosis and numerous lysosomes associated with autophagic vacuoles showing signs of apoptosis. In conclusion, this study confirms that the ultrastructural characteristics of pESCs differ depending on their origin. These ultrastructural characteristics might be useful in biomedical research using pESCs, leading to new insights regarding regenerative medicine and tissue repair.

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Section: Methodsmentioning

confidence: 99%

Ultrastructural comparison of porcine putative embryonic stem cells derived by <i>in vitro</i> fertilization and somatic cell nuclear transfer

Yoo

Kim

Hwang

et al. 2016

Journal of Reproduction and Development

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“…Additionally, mitochondrial morphology is regarded as a novel factor for determining pluripotent states [ 28 ]. Consistent with this, our recent work has also demonstrated that porcine ESCs derived from porcine clone blastocysts using iPSCs as donors (iPS-NT ESCs)—which have relatively more naïve pluripotent features—have a more immature form of the mitochondrial organelle shape with under-developed cristae than other origins regarded as the primed state [ 29 ]. This result suggested that a difference in mitochondrial dynamics is present not only between ESCs and differentiated cells but also between pluripotent stem cells of different pluripotent states (naïve vs. primed).…”

Section: Embryonic Stem Cells and Apoptosismentioning

confidence: 57%

Apoptosis in Porcine Pluripotent Cells: From ICM to iPSCs

Kim

Hyun

2016

IJMS

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Abstract:Pigs have great potential to provide preclinical models for human disease in translational research because of their similarities with humans. In this regard, porcine pluripotent cells, which are able to differentiate into cells of all three primary germ layers, might be a suitable animal model for further development of regenerative medicine. Here, we describe the current state of knowledge on apoptosis in pluripotent cells including inner cell mass (ICM), epiblast, embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs). Information is focused on the apoptotic phenomenon in pluripotency, maintenance, and differentiation of pluripotent stem cells and reprogramming of somatic cells in pigs. Additionally, this review examines the multiple roles of apoptosis and summarizes recent progress in porcine pluripotent cells.

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“…Regardless of the clonal variation in some stem cell properties, the established piPSC-ntESC lines could all form teratomas in SCID mice. Kim et al [40] reported putative porcine ESC lines derived from cloned blastocysts reconstructed with piPSCs. Those piPSCs were generated using plasmid vectors harboring four factors: Oct4, Klf4, Sox2, and cMyc [41].…”

Section: Discussionmentioning

confidence: 99%

Establishment of porcine nuclear transfer-derived embryonic stem cells using induced pluripotent stem cells as donor nuclei

Haraguchi

Dang‐Nguyen

Wells

et al. 2020

Journal of Reproduction and Development

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We investigated whether sequential reprogramming via porcine induced pluripotent stem cells (piPSCs) or exposure to oocyte cytoplasm following nuclear transfer could generate nuclear transfer-derived ESCs (piPSCs-ntESCs). Nuclear transfer embryos were reconstructed with piPSCs possessing a ZsGreen fluorescent marker for expression of exogenous Nanog and Lin28. Reconstructed oocytes developed to morphologically normal 8-cell/morulae (35/93, 37.6%) and blastocysts (12/93, 12.9%). Although most green fluorescent protein-positive blastocysts showed efficient outgrowth (8/10, 80%), none formed primary colonies and all cultures degenerated. Conversely, 15% of fluorescent positive 8-cell/morula stage embryos showed outgrowth (6/40), with three forming primary colonies (7.5%). All three were expanded and maintained as piPSC-ntESC lines. These cell lines expressed stem cell marker genes and proteins. Despite inactivation of one X chromosome, all piPSC-ntESC lines formed teratomas comprising derivatives from all three embryonic germ layers. Strong SSEA1, 3, and 4 expression was detected at the 8-cell/morula stage in embryos reconstructed from both piPSCs and porcine embryonic fibroblasts (PEFs). SSEA3 was notably absent from IVF controls at pre-implantation embryo stages. Finally, we attempted to establish ntESCs from 8-cell/morulae reconstructed with PEFs using the same culture conditions as those for piPSC-ntESC derivation. Although eight primary colonies arose from 107 embryos (7.5%), they all degenerated after the first passage culture. Early and sustained expression of key reprogramming regulatory factors may be critical for pluripotent stem cell derivation to derive piPSC-ntESCs from 8-cell/morula stages, while the expression of SSEAs may be involved in the initial stem cell colony formation phases.

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Putative embryonic stem cells derived from porcine cloned blastocysts using induced pluripotent stem cells as donors

Cited by 20 publications

References 80 publications

Ultrastructural comparison of porcine putative embryonic stem cells derived by <i>in vitro</i> fertilization and somatic cell nuclear transfer

Ultrastructural comparison of porcine putative embryonic stem cells derived by <i>in vitro</i> fertilization and somatic cell nuclear transfer

Apoptosis in Porcine Pluripotent Cells: From ICM to iPSCs

Establishment of porcine nuclear transfer-derived embryonic stem cells using induced pluripotent stem cells as donor nuclei

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