Pluripotent and Metabolic Features of Two Types of Porcine iPSCs Derived from Defined Mouse and Human ES Cell Culture Conditions

The ability to generate germ cells from pluripotent stem cells (PSCs) is valuable for human regenerative medicine and animal breeding. Germ cell-like cells (GCLCs) have been differentiated from mouse and human PSCs, but not from porcine PSCs, which are considered an ideal model for stem cell applications. Here, we developed a defined culture system for the induction of primordial germ cell-like cells (PGCLCs) from porcine induced PSCs (piPSCs). The identity of the PGCLCs was characterized by observing cell morphology, detecting germ cell marker gene expression and evaluating epigenetic properties. PGCLCs could further differentiate into spermatogonial stem cell-like cells (SSCLCs) in vitro. Importantly, meiosis occurred during SSCLC induction. Xenotransplantation of GCLCs into seminiferous tubules of infertile immunodeficient mice resulted in immunohistochemically identifiable germ cells in vivo. Overall, our study provides a feasible strategy for directing piPSCs to the germ cell fate and lays a foundation for exploring germ cell development mechanisms.

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“…The iPSCs were maintained in 2i plus LIF medium20. Similar to mouse PSCs, piPSCs showed a compact and dome-like colony morphology (Fig.…”

Section: Resultsmentioning

confidence: 99%

Induction of Germ Cell-like Cells from Porcine Induced Pluripotent Stem Cells

Wang

Xiang

Zhang

et al. 2016

Sci Rep

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“…Therefore, X-chromosome activation is considered to be a key indicator that a pluripotent cell is in a naive or primed state [155] . For porcine iPSCs, three cell lines generated by the expression of transcription factors OCT4, KLF4 and C-MYC showed naive-like iPSC properties with an activated X-chromosome, as well as high embryonic chimera incorporation efficiency [97,126,130] . In addition, Haraguchi and colleagues successfully established a unique cell line derived from ICM of porcine embryo that exhibited LIF-dependency but not bFGFdependency, and could be considered as naive-state cells as mouse ESCs; however, the X-chromosome status in this cell line was XaXi [45] .…”

Section: X-chromosome Activationmentioning

confidence: 99%

“…Relative to the embryonic stem cell lines, studies on iPSCs in large animals has made more progress, especially for pigs. However, there are still some difficulties, cell propagation is dependent on exogenous genes and cannot produce germline chimeric offspring [59,127,130,164,165] . These problems need to be solved urgently for the study of PSCs in large animals.…”

Section: Challenges and Prospectsmentioning

confidence: 99%

Porcine pluripotent stem cells: progress, challenges and prospects

Han¹,

Miao²,

Hua³

et al. 2019

Front. Agr. Sci. Eng.

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Pluripotent stem cells (PSCs) are characterized by their capacity for high self-renewal and multiple differentiation potential and include embryonic stem cells, embryonic germ cells and induced PSCs. PSCs provide a very suitable model for the studies of human diseases, drugs screening, regenerative medicine and developmental biology research. Pigs are considered as an ideal model for preclinical development of human xenotransplantation, therapeutic approaches and regenerative medicine because of their size and physiological similarity to humans. However, lack of knowledge about the derivation, characterization and pluripotency mechanisms of porcine PSCs hinders progress in these biotechnologies. In this review, we discuss the latest progress on porcine PSCs generation, evaluation criteria for pluripotency, the scientific and technical questions arising from these studies. We also introduce our perspectives on porcine PSC research, in the hope of providing new ideas for generating naive porcine PSCs and animal breeding.

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“…Therefore, a key indication of complete reprogramming is the ability of iPSCs to maintain PSCs characteristics in absence of reprogramming gene expression. Nevertheless, the continuous expression of reprogramming factors in iPSCs does not seem to interfere with self-renewal and pluripotency (West et al 2010; Zhang et al 2015). Similar to ESCs, iPSCs can be maintained under different pluripotency states, ranging from naïve to region-selective primed cells.…”

Section: Induced Pluripotencymentioning

confidence: 99%

“…Interestingly, many of the studies summarized in Table 1 describing derivation of naïve state livestock iPSCs do not evaluate X-chromosome reactivation in female cells. Only three studies (Whitworth et al 2014a; Zhang et al 2014, 2015) evaluated H3K27me3 of X-chromosome by immunostaining showing promising results. X-chromosome reactivation is an important feature of naïve state pluripotency and its detection can add valuable information to the epigenetic profile of reprogrammed somatic cells.…”

Section: Pscs In Livestock Species and Applications For Generation Ofmentioning

confidence: 99%

Pluripotent stem cells and livestock genetic engineering

Soto

Ross

2016

Transgenic Res

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The unlimited proliferative ability and capacity to contribute to germline chimeras make pluripotent embryonic stem cells (ESCs) perfect candidates for complex genetic engineering. The utility of ESCs is best exemplified by the numerous genetic models that have been developed in mice, for which such cells are readily available. However, the traditional systems for mouse genetic engineering may not be practical for livestock species, as it requires several generations of mating and selection in order to establish homozygous founders. Nevertheless, the self-renewal and pluripotent characteristics of ESCs could provide advantages for livestock genetic engineering such as ease of genetic manipulation and improved efficiency of cloning by nuclear transplantation. These advantages have resulted in many attempts to isolate livestock ESCs, yet it has been generally concluded that the culture conditions tested so far are not supportive of livestock ESCs self-renewal and proliferation. In contrast, there are numerous reports of derivation of livestock induced pluripotent stem cells (iPSCs), with demonstrated capacity for long term proliferation and in vivo pluripotency, as indicated by teratoma formation assay. However, to what extent these iPSCs represent fully reprogrammed PSCs remains controversial, as most livestock iPSCs depend on continuous expression of reprogramming factors. Moreover, germline chimerism has not been robustly demonstrated, with only one successful report with very low efficiency. Therefore, even 34 years after derivation of mouse ESCs and their extensive use in the generation of genetic models, the livestock genetic engineering field can stand to gain enormously from continued investigations into the derivation and application of ESCs and iPSCs.

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Pluripotent and Metabolic Features of Two Types of Porcine iPSCs Derived from Defined Mouse and Human ES Cell Culture Conditions

Cited by 30 publications

References 44 publications

Induction of Germ Cell-like Cells from Porcine Induced Pluripotent Stem Cells

Induction of Germ Cell-like Cells from Porcine Induced Pluripotent Stem Cells

Porcine pluripotent stem cells: progress, challenges and prospects

Pluripotent stem cells and livestock genetic engineering

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