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Performance of Transgenic Three-Genes Expressed Pig for Alzheimer's Disease Model
Yeo-Jin Son,Seung-Eun Lee,Youngsok Choi,Mi-Ryung Park,Hyun-Sok Hong,Min-Young Shin,Yun-Gwi Park,Sang-Gi Jeong,Young-Ho Kim,Gi-sun Im,Eun-Young Kim,Se-Pill Park 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
Alzheimer's disease (AD) has caused by expression of amyloid precursor protein (APP), Tau and presenilin (PS) as known as plaques and tangle accumulation. AD transgenic porcine model is necessary for preclinical testing of therapeutic agent because of similar metabolic system between porcine and human. The objective of study was to generate AD transgenic pig by somatic cell nuclear transfer (SCNT) with multi-cistronic vector system. AD multi-cistronic vector was 6 well-known mutation on 3 AD related genes, hAPP (K670N/M671L, I716V, V717I), hTau (P301L) and hPS1 (M146V, L286P). Establishment of AD transgenic cell lines was used from Jeju black pig ear fibroblast cells (JB-PEFAD) with the AD multi-cistronic vector. The JB-PEFAD cell was confirmed on mRNA expression, protein synthesis of hAPP, hTau and hPS1 and identification of integration and karyotype. Although fusion rate was no difference in SCNT with JB-PEF AD (SCNTAD) embryos, cleavage and blastocyst formation rates were slightly lower than in SCNT with non-transgenic JB-PEF (SCNTnon-TG). Individual SCNTAD blastocysts were detected hAPP, hTau and hPS1 genomic integration which showed 93.2% (n=30) efficiency in genomic DNA (gDNA) level. It will give us a possibility to develop porcine animal model for AD study in the future.
Establishment of Induced Pluripotent Stem Cells using Jeju Black Pig Fibroblast Cells
Yeo-Jin Son,Seung-Eun Lee,Eun-Young Kim,Se-Pill Park 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & Developmental Biology(Supplement) Vol.41 No.2
Stem cells can be differentiated into numerous cell types and then can be used to research intractable disease and regenerative medicine. Induced pluripotent stem cells (iPSCs) are produced through an artificial manipulation in vitro, and have the advantage of being to customized to patient. In this study, we used the Sendai virus to produce porcine iPSC (p-iPSC) from Jeju Black Pig (JBP) cells. JBP cell was infected for 24 h with Sendai virus (1×104 cells/mL). One week after infection, they were spread onto mouse embryonic fibroblast (MEF) feeder cell layer. When we used JBP #4007 2nd passage cells for iPSC generation, primary colony was appeared between 3~4 weeks. The first and second colony were dissociated by treatment with trypsin and hereafter formed colony were mechanically dissociated by insulin syringe. During culture of the JBP-iPSC, some colonies were stained by alkaline phosphatase (AP) staining kit for confirming expression of the pluripotency marker. The stem cell markers as undifferentiation state, Oct4, Sox2, SSEA-1 and Nanog, were stained using JBP-iPSC at 5 passage. The JBP-iPSC colony have been maintaining in nowaday 14 passages, and the AP and immunostain were expressed. These results demonstrate that p-iPSC can derive from Jeju Black Pig cell using Sendai virus, and it will be used to stem cell research for animal disease model.
Yeo Jin Lee,Young Min Son,Min Jeong Gu,Ki-Duk Song,Sung-Moo Park,Hyo Jin Song,Jae Sung Kang,Jong Soo Woo,Jee Hyung Jung,Deok-Chun Yang,Seung Hyun Han,Cheol-Heui Yun 고려인삼학회 2015 Journal of Ginseng Research Vol.39 No.1
Background: Panax ginseng (i.e., ginseng) root is extensively used in traditional oriental medicine. It is a modern pharmaceutical reagent for preventing various human diseases such as cancer. Ginsenosidesd-the major active components of ginsengdexhibit immunomodulatory effects. However, the mechanism and function underlying such effects are not fully elucidated, especially in human monocytes and dendritic cells (DCs). Methods: We investigated the immunomodulatory effect of ginsenosides from Panax ginseng root on CD14⁺ monocytes purified from human adult peripheral blood mononuclear cells (PBMCs) and on their differentiation into DCs that affect CD4⁺ T cell activity. Results: After treatment with ginsenoside fractions, monocyte levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10 increased through phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK), but not p38 mitogen-activated protein kinase (MAPK). After treatment with ginsenoside fractions, TNF-α production and phosphorylation of ERK1/2 and JNK decreased in lipopolysaccharide (LPS)-sensitized monocytes.We confirmed that DCs derived from CD14⁺ monocytes in the presence of ginsenoside fractions (Gin-DCs) contained decreased levels of the costimulatory molecules CD80 and CD86. The expression of these costimulatory molecules decreased in LPS-treated DCs exposed to ginsenoside fractions, compared to their expression in LPS-treated DCs in the absence of ginsenoside fractions. Furthermore, LPS-treated Gin-DCs could not induce proliferation and interferon gamma (IFN-γ) production by CD4⁺ T cells with the coculture of Gin-DCs with CD4⁺ T cells. Conclusion: These results suggest that ginsenoside fractions from the ginseng root suppress cytokine production and maturation of LPS-treated DCs and downregulate CD4⁺ T cells.
Performance of Transgenic Three-Genes Expressed Pig for Alzheimer's Disease Model
Yeo-Jin Son,Seung-Eun Lee,Youngsok Choi,Mi-Ryung Park,Hyun-Sok Hong,Min-Young Shin,Yun-Gwi Park,Sang-Gi Jeong,Young-Ho Kim,Gi-sun Im,Eun-Young Kim,Se-Pill Park 한국동물생명공학회(구 한국동물번식학회) 2016 발생공학 국제심포지엄 및 학술대회 Vol.2016 No.10
Yeo Jin Lee,Young Min Son,Min Jeong Gu,Ki-Duk Song,Sung-Moo Park,Hyo Jin Song,Jae Sung Kang,Jong Soo Woo,Jee Hyung Jung,Deok-Chun Yang,Seung Hyun Han,Cheol-Heui Yun 고려인삼학회 2015 Journal of Ginseng Research Vol.39 No.3
Background: Panax ginseng (i.e., ginseng) root is extensively used in traditional oriental medicine. It is a modern pharmaceutical reagent for preventing various human diseases such as cancer. Ginsenosidesdthe major active components of ginsengdexhibit immunomodulatory effects. However, the mechanism and function underlying such effects are not fully elucidated, especially in human monocytes and dendritic cells (DCs). Methods: We investigated the immunomodulatory effect of ginsenosides from Panax ginseng root on CD14⁺ monocytes purified from human adult peripheral blood mononuclear cells (PBMCs) and on their differentiation into DCs that affect CD4⁺ T cell activity. Results: After treatment with ginsenoside fractions, monocyte levels of tumor necrosis factor (TNF)-a, interleukin (IL)-6, and IL-10 increased through phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK), but not p38 mitogen-activated protein kinase (MAPK). After treatment with ginsenoside fractions, TNF-α production and phosphorylation of ERK1/2 and JNK decreased in lipopolysaccharide (LPS)-sensitized monocytes.We confirmed that DCs derived from CD14⁺ monocytes in the presence of ginsenoside fractions (Gin-DCs) contained decreased levels of the costimulatory molecules CD80 and CD86. The expression of these costimulatory molecules decreased in LPS-treated DCs exposed to ginsenoside fractions, compared to their expression in LPS-treated DCs in the absence of ginsenoside fractions. Furthermore, LPS-treated Gin-DCs could not induce proliferation and interferon gamma (IFN-γ) production by CD4⁺ T cells with the coculture of Gin-DCs with CD4⁺ T cells. Conclusion: These results suggest that ginsenoside fractions from the ginseng root suppress cytokine production and maturation of LPS-treated DCs and downregulate CD4⁺ T cells.
Yeo-Jin Son,Seung-Eun Lee,Min-Young Shin,Yun-Gwi Park,Sang-Gi Jeong,Eun-Young Kim,Se-Pill Park 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
Somatic cell nuclear transfer (SCNT) technique is a key point of producing transgenic animal disease models. During in vitro production of SCNT embryo, the quality of matured oocytes are one of the important factors that regulate embryo developmental capacity. In preliminary test, we confirmed the effect of fibroblast growth factor 10 (FGF10) on porcine oocyte maturation. In this study, we investigated the developmental potential of SCNT embryos treated with the 10 ng/ml FGF10 (10 F) during in vitro maturation of recipient oocytes. The polar body emission rate was significantly higher in the 10 F treated group than control group. After SCNT, although the rate of fusion was no significant difference, the rate of cleavage and blastocyst formation was significantly increased in the 10 F treated group (p<0.05). In 10 F treated group, the total cell number was increased and the percentage of apoptotic cell was decreased in the blastocyst stage at day 7 (p<0.1). The transcription level of apoptosis relative gene, Casp3 was significantly decreased, while anti-apoptosis gene BCL2l1 was increased in the 10 F treated group compared to control group. The 10 F treated group was highly expressed the reprogramming related genes, Sox2 and POU5f1. Also, the first cleaving time was more faster and the percentage of cell block was significantly lower in 10 F treated group than in control group. In this study, we confirmed that 10 ng/ml FGF10 has effect on enhance the oocyte maturation and developmental capacity. These results demonstrate that FGF10 treatment can be used for in vitro development of porcine SCNT embryos and subsequent production of transgenic animal model.