Pluripotent stem cells could self-renew and differentiate into various cells. In particular, porcine pluripotent stem cells are useful for preclinical therapy, transgenic animals, and agricultural usage. These stem cells have naïve and primed pluripotent states. Naïve pluripotent stem cells represented by mouse embryonic stem cells form chimeras after blastocyst injection. Primed pluripotent stem cells represented by mouse epiblast stem cells and human embryonic stem cells. They could not produce chimeras after blastocyst injection. Populations of embryonic stem cells are not homogenous; therefore, reporter systems are used to clarify the status of stem cells and to isolate the cells. For this reason, studies of the OCT4 reporter system have been conducted for decades. This review will discuss the naïve and primed pluripotent states and recent progress in the development of porcine OCT4 reporter systems.

As a preclinical study, many researchers have been attempted to convert the porcine PSCs into several differentiated cells with transplantation of the differentiated cells into the pigs. Here, we attempted to derive neuronal progenitor cells from pig embryonic germ cells (EGCs). As a result, neuronal progenitor cells could be derived directly from pig embryonic germ cells through the serum-free floating culture of EB-like aggregates (SFEB) method. Treating retinoic acid was more efficient for inducing neuronal lineages from EGCs rather than inhibiting SMAD signaling. The differentiated cells expressed neuronal markers such as PAX6, NESTIN, and SOX1 as determined by qRT-PCR and immunostaining. These data indicated that pig EGCs could provide valid models for human therapy. Finally, it is suggested that developing transgenic pig for disease models as well as differentiation methods will provide basic preclinical data for human regenerative medicine and lead to the success of stem cell therapy.

Stem cells are progenitor cells that are capable of self-renewal and differentiation into various cells. Especially, pluripotent stem cells (PSCs) have in vivo and in vitro differentiation capacity into three germ layers and can proliferate infinitely. The differentiation ability of PSCs can be applied for regenerative medicine and tissue engineering. In domestic animals, their PSCs have a potential for preclinical therapy as well as the production of transgenic animals and agricultural usage such as cultured meat. Among several domestic animals, a pig is considered as an ideal model for biomedical and agricultural purposes mentioned above. In this reason, studies for pig PSCs including embryonic stem cells (ESCs), embryonic germ cells (EGCs) and induced pluripotent stem cells (iPSCs) have been conducted for decades. Therefore, this review will discuss the history of PSCs derived from various origins and recent progress in pig PSC research field.

Because of the physiological and immunological similarities between pigs and humans, porcine embryonic stem cells (ESCs) have been identified as important candidates in preliminary studies on human disease. A comparative understanding of pig ESCs with the human is required to achieve these goals. To gain insights into pig stem cells, the transcriptome of pig ES-like cells were compared with pig preimplantation embryos and human/mouse pluripotent stem cells by RNA-seq analysis. As a result, pig stem cells were more similar to late epiblasts of pig preimplantation embryos than early ICM as revealed by transcriptome analysis, suggesting that pig stem cells are in a developmentally primed state. Moreover, the physiological and biological functions of pig ESCs were more similar to those of human PSCs than to those of mouse PSCs, as determined by direct differentiation and GO/KEGG term analysis. Overall, our data indicate that pig ESCs are in a primed pluripotent state resembling human PSCs. Our findings will facilitate both the development of large animal models for human stem cell therapy and the generation of pluripotent stem cells from other domestic animals for agricultural use. This work was supported by the Korea Institute of Planning and Evaluation for Technology in food, agriculture, forestry, and fisheries (IPET) through the Development of High Value-Added Food Technology Program funded by the Ministry of Agriculture, Food, and Rural Affairs (MAFRA; 118042-03-1-HD020), and partially supported by the grants from the Agenda Program of Rural Development Administration, Republic of Korea (No. PJ01362402)

Because the pig is a valuable candidate for a preclinical model of human cell therapy as well as an important food source, understanding a physiology of pig myogenic progenitors such as skeletal muscle satellite cells and myoblasts is required for cure of muscular diseases and improvement of meat production. For these reasons, we tried to isolate and culture the pig progenitor cells from skeletal muscle. Pig satellite cells, known as muscle stem cells, were isolated from biceps femoris of neonatal pigs by enzymatic digestion method. Muscle satellite cells are quiescent in uninjured muscle. Upon injury, they are activated into proliferating state, known as myoblasts, by growth factors and, in turn, differentiated forward to myocytes and myotubes. To trigger proliferation in vitro, the isolated satellite cells were cultured with epidermal growth factor (EGF) and dexamethasone (BMP4 inhibitor). As a result, the pig satellite cells were efficiently converted into proliferating myoblasts and stably maintained over an extended period. The myoblasts were confirmed by markers of PAX7, MYF5, and MYOD1. Our finding showed that modulating EGF and BMP4 signaling are essential for maintaining muscle stem cells. This culture method could be applied for a production of cultured meat and further basic research of muscle development. This work was supported by the Korea Institute of Planning and Evaluation for Technology in food, agriculture, forestry, and fisheries (IPET) through the Development of High Value-Added Food Technology Program funded by the Ministry of Agriculture, Food, and Rural Affairs (MAFRA; 118042-03-1-HD020).

The transcription factor POU5F1, also known as OCT4 plays critical roles in maintaining pluripotency during early mammalian embryonic development and in embryonic stem cells. It is important to establish an OCT4 promoter region-based reporter system to study pluripotency. However, there is still a lack of information about the porcine OCT4 upstream region. To improve our understanding of the porcine OCT4 regulatory region, we identified conserved regions in the porcine OCT4 promoter upstream region by sequence-based comparative analysis using various mammalian genome sequences. The similarity of nucleotide sequences in the 5' upstream region was low among mammalian species. However, the OCT4 promoter and four regulatory regions, including distal and proximal enhancer elements, had high similarity. The putative transcription factor binding sites in the Oct4 5' upstream region nucleotide sequences from mice and pigs also differed. Some of these genes are related to pluripotency, and further research will allow us to better understand the differences in species-specific pluripotency. Next, a functional analysis of the porcine OCT4 promoter region was conducted. Luciferase reporter assay results indicated that the porcine OCT4 distal enhancer and proximal enhancer were highly activated in mouse embryonic stem cells and embryonic carcinoma cells, respectively. Similar to OCT4 upstream-based reporter systems derived from other species, the porcine OCT4 upstream region-based reporter constructs showed exclusive expression patterns depending on the state of pluripotency. This work provides basic information about the porcine OCT4 upstream region and various porcine OCT4 fluorescence reporter constructs, which can be applied to study species-specific pluripotency in early embryo development and the establishment of embryonic stem cells in pigs. This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education (NRF-2017R1D1A1B03032256).

The impact of sodium hydroxide, which is one of chemicals of clean in place (CIP) for removing membrane fouling, on the PVDF membrane is reviewed with respect to physical/chemical structural change, the permeability affected therefrom. Based on the cleaning concentration applied in membrane water treatment facilities, 10% of accumulated defluorination was confirmed up to 166g.hr/L which reflects the exposure time. However, membrane resistance was confirmed to be reduced by about 10%. Through FT-IR and EDS analysis, reduction of F and change of are confirmed as factors that affect the permeability of membrane. Membrane resistance, which affects permeability, is affected by loss of additives for hydrophilicity, rather than defluorination of PVDF material. Therefore, in order to check membrane degradation degree, an accelerated test by NaOH was carried out, loss of additives was confirmed, and then PVDF inherent characteristic was observed.

포식성이리응애류는 섭식행동이 왕성한 천적군으로 알려져 있으며, 다수의 종들이 상업적으로 개발되고 있다. 이에 우리는 국내 토착 포식성이리응애의 이용도를 탐색하였다. 2012년까지 국내에서 서식이 보고된 포식성이리응애류은 47종을 대상으로 구글 학술검색을 이용하여 논문수를 조사한 결과, 사막이리응애가 3,560개로 가장 많은 수가 검색되었다. 천개 이상의 논문이 검색된 종은 서양이리응애(3,160개), 나팔이리응애(1,590개), 순이리응애(1,510개), 대중이리응애(1,260개), 긴털이리응애(1,190개), 동양이리응애 (1,020개)이었다. 백개 이상의 논문이 검색된 종은 긴꼬리이리응애(314개), 알락이리응애(109개), 엷은줄이리응애(345개), 북방이리응애(143개), 꽃무늬이리응애(200개) 등 5종이었다. 이 중 전세계적으로 상품화가 보고된 종은 사막이리응애, 서양이리응애, 나팔이리응애이다. 국내에 서식이 보고되지 않았으나, 국내외 농업현장에서 가장 많이 사용되는 지중해이리응애는 5,060개가 보고되었다. 긴꼬리이리응애는 국내에서 47종의 식물에서 서식하여 가장 많은 식물 종에서 발견된 포식성이리응애로 농업해충인 점박이응애, 총채벌레류, 가루이류를 포식하며, 농업해충 외에 농작물해충이 아닌 긴털가루응애, 설탕응애, 꽃가루 등의 섭식을 확인하여 앞으로 국내에서 농업해충을 방제하기 위한 천적으로 활용여부에 대한 적극적인 검토가 요구된다.

X-chromosome inactivation is one of the most complex events observed in early embryo developments. The epigenetic changes occurred in female X-chromosome is essential to compensate dosages of X-linked genes between males and females. Because of the relevance of the epigenetic process to the normal embryo developments and stem cell studies, X-chromosome inactivation has been focused intensively for last 10 years. Initiation and regulation of the process is managed by diverse factors. Especially, proteins and non-coding RNAs encoded in X-chromosome inactivation center, and a couple of transcription factors have been reported to regulate the event. In this review, we introduce the reported factors, and how they regulate epigenetic inactivation of X-chromosomes.

It is still challenging to establish pESCs due to differences in the genetic backgrounds of mouse, human, and pig. So it is required to find pig specific pluripotency markers and cellular signaling. In this experiments, doxycycline-inducible vectors carrying OCT4, SOX2, NANOG, KLF4 and MYC known as reprogramming factors, were infected into pig stem cells for analyzing gene expression pattern. When cultured without doxycycline, pig stem cells were stably maintained in bFGF supplemented media. However, when treated with doxycycline, pig stem cells lost alkaline phosphatase activity and were differentiated within two weeks. And then, we investigated the expression of genes related to pluripotency in doxycycline-treated pig stem cells by using qRT-PCR. The qRT-PCR data revealed that expression of OCT4, CDH1 and FUT4 were significantly increased by OCT4 overexpression and OCT4 and FUT4 were also upregulated in SOX2-infected group. When infected with combination of two factors including OCT4 or SOX2, some groups could stably maintain at LIF supplemented media, having alkaline phosphatase activity. Given these data, although ectopic gene expression induced differentiation in pig stem cells, ectopic expression of OCT4 and SOX2 could upregulate pluripotent genes and overexpreession of two factors help pig stem cells adapt LIF-contained media. This study could improve understanding of pluripotent networks as well as aid in establishing bona fide pluripotent stem cells in pig.

Various studies have forwarded an outstanding wastewater effluent treatment systems toward securing sustainable supply of water sources. In this paper, a broad overview of the performance of MF membrane as pretreatment option for wastewater reuse will be presented based on the literature survey and experiments conducted over the wastewater reuse pilot plant. The pilot plant was operated with a continuous data acquisition for about 300days under various chemical enhanced backwash (CEB) system with subsequent treated water quality analysis. Accordingly, assessment of the effluent revealed that the pretreated water is suitable enough to be used as an input for Reverse Osmosis (RO) unit and significant effect of CEB and concentration of NaOCl is also conceived from the analysis. Moreover, it's also observed that the application of various CEB condition over long operational hours induced a constant declination of overall performance of MF membrane.

The effects of dissolved inorganic and organic matter in seawater and the characteristics of fouling on the membrane surface were investigated within membrane distillation (MD) process. The changes of the membrane flux of PE and PVDF hollow fiber membranes under natural and synthetic seawater were compared with given variances of temperature. The flux of both membranes under the synthetic seawater, without any organic matter, were higher than that of the natural seawater, indicating the organic fouling on the membrane surface. The surface of the membrane was analyzed using scanning electron microscope (SEM) to examine the fouling. The experiment with organics has shown the formation of thin film over the membrane surface, while the experiment with inorganics has shown only the formation of inorganic crystals. The results indicated the organic matter as the major foulants and that the organics affected the formation of the crystals. Permeate water conductivity of all conditions verified the quality of the water to be better if not similar to that of RO.

MicroRNAs (miRNAs) are approximately 22 nucleotides of small noncoding RNAs that control gene expression at the posttranscriptional level through translational inhibition and destabilization of their target mRNAs. The miRNAs are phylogenetically conserved and have been shown to be instrumental in a wide variety of key biological processes including cell cycle regulation, apoptosis, metabolism, imprinting, and differentiation. Recently, a paper has shown that expression of the miRNA-302/367 cluster expressed abundantly in mouse and human embryonic stem cells (ESCs) can directly reprogram mouse and human somatic cells to induced pluripotent stem cells (iPSCs) efficiently in the absence of any of the four factors, Oct4, Sox2, c-Myc, and Klf4. To apply this efficient method to porcine, we analyzed porcine genomic sequence containing predicted porcine miRNA-302/367 cluster through ENSEMBL database, generated a non-replicative episomal vector system including miRNA-302/367 cluster originated from porcine embryonic fibro-blasts (PEF), and tried to make porcine iPSCs by transfection of the miRNA-302/367 cluster. Colonies expressing EGFP and forming compact shape were found, but they were not established as iPSC lines. Our data in this study show that pig miRNA-302/367 cluster could not satisfy requirement of PEF reprogramming conditions for pluripotency. To make pig iPSC lines by miRNA, further studies on the role of miRNAs in pluripotency and new trials of transfection with conventional reprogramming factors are needed.

Yellow clay dispersion has been applied to minimize fisheries impact by the red tide Cochlodinium polykrikoides blooms in Korean coasts since 1995. The present preliminary study documents the effect of yellow clay on Korean rockfish, Sebastes schlegelii, in terms of oxygen consumption rate (OCR). The OCR in the low clay suspension (0.05 and 0.23 %, w/w) showed normal level compared to the control. In contrast, the OCR for each one of three replicates in the high clay suspension (1.16 and 5.58 %, w/w) was not returned to the previous level that clay was not treated, indicating that high clay suspension (≥1.16%, w/w) might give negative effect on Korean rockfish. Overall, this result suggests that field application of clay to control Harmful Algal Blooms (HABs) may not give impact on Korean rockfish once the clay is dispersed in a low concentration (≤0.23%). In order to understand the changes of OCR in the repeated exposure to clay, it is required to do further studies on the changes of OCR when the fish is exposed to clay repeatedly after recovery in the normal seawater.

Pig embryonic stem cells (ESC) has been suggested to become important animal model for therapeutic cloning using embryonic stem cells derived by somatic cell nuclear transfer (SCNT). However, the quality of cloned embryo and derivation rate of cloned blastocyst has been presented limits for derivation of cloned embryonic stem cell. In this study, we have tried to overcome these problems by aggregating porcine embryos. Zonafree reconstructed SCNT Embryos were cultured in micro-wells singularly (non-aggregated group) or as aggregates of three (aggregated groups) at the four cell stage. Embryo quality of the cloned embryos and attachment on feeder layer rate significantly increased in the aggregates. The aggregation of pig SCNT embryos at the four-cell stage can be a useful technique for improving the quality of pig cloned blastocyst and improvement in the percentage of attachment on the feeder layer of cloned embryos. * This work was supported by the BioGreen 21 Program (PJ0081382011), Rural Development Administration, Republic of Korea.

A recent study has reported that pluripotent stem cells can be categorized according to their pluripotent state. The first is a “naïve” state, which is characterized by small, round or dome-shaped colony morphologies, LIF and BMP4 signaling pathways and two active X chromosomes in female; mouse ES cells (mESCs) represent this type. A second “primed” state has also been described and is possible in mouse epiblast stem cells (mEpiSCs) or human ES cells (hESCs). These primed state pluripotent stem cells display flattened monolayer colony morphologies, FGF and Nodal/Activin signaling pathways and X chromosome inactivation in female. It has been suggested that, as a non-permissive species, the porcine species undergoes reprogramming into a primed state during the establishment of pluripotent stem cell lines. Meanwhile, a few studies have reported that primed pluripotent stem cell lines could be reverted to a naïve pluripotent state using various exogenous factors including GSK3β and MEK inhibitors, LIF, hypoxic conditions and up-regulation of Oct3 or klf4. Therefore, the purpose of this study was to investigate whether a LIF-dependent naïve pluripotent stem cell line could be derived from porcine embryonic fibroblasts(PEFs) via doxycycline (dox)-inducible reprogramming factors and LIF. In this study, we have been able to successfully induce PEFs into a LIF-dependent naïve pluripotent-like cell line showing a mESC-like morphology and the expression of pluripotent markers. Our results suggest the possibility of reprogramming to naive pluripotent- like stem cells from PEFs in porcine species. * This work was supported by the BioGreen 21 Program (PJ0081382011), Rural Development Administration, Republic of Korea.