긴볼레기말의 항고지혈증 효과에 관한 연구 : Triton WR-1339 주사에 의한 고지혈증 유발 생쥐의 간세포내 지방 축적 감소를 중심으로 Based on the Decreasing Effect of Lipid Accumulation in Hepatocyte of Murine with Hyperlipidemia induced by Triton WR-1339

박인식,안상현,정재만,강윤호,이해풍,서귀문,홍용기,김호현,김진택 동국대학교 한의학연구소 1999 東國韓醫學硏究所論文集 Vol.7 No.2

본 연구는 긴볼레기말 추출물의 항고지혈증 효과를 조사하기 위해 ICR 생쥐에 TritonWR-1339(TX) 복강주사로 인위적인 고지혈증을 유발시킨 후 긴볼레기말 추출물(30㎎/㎏)를 복강주사하여 시간의 경과에 따른 간세포내에서의 지방 축적 변화를 조직화학적으로 관찰하였다. TX 주사후 그물구조의 세포질출 가진 간세포가 간엽 전체에서 관찰되었고, 일부 간소엽에서는 간세포 손상으로 인한 간세포판 소실이 나타났다. 또한 간세포내 지방축척도 증가하여 전체 간소엽의 간세포에서 지방의 과출현을 확인 할 수 있었고, 지방의 크기도 대조군에 비해 증가된 것으로 관찰되었다. 그러나 긴볼레기말 추출물 주사군에서는 그물구조의 세포질을 가진 간세포의 수가 TX 주사군에 비해 감소되었고, 대부분의 간소엽에서 정상적인 간세포판의 배열을 확인할 수 있었다. 간세포내의 지방 축적과 크기도 감소된 경향으로 관찰되었다. 이상의 결과로 볼 때 해조류 긴볼레기말 추출물은 고지혈증이 유발된 생쥐 간세포 내에서의 과도한 지방축적을 감소시키는 항고지혈증 효과를 하는 것으로 사료된다. Hepatic tissues of ICR mouse were intraperitoneally injeced with Colpomenia bullosa(CB) Extract after Triton WR-1339(TX) injection were observed to investigate the antihyperlipermic effect of CB extract for hyperlipidemic hepatic tissue caused by destruction of lipid metabolism. The hepatic tissues were obtained at hour-24, 48, and 72 after TX injection with CB extract treatment. And then these specimen were fixed in 10% neutral buffer solution and were cryocut. The tissue stained by H&E for general morphology and sudan black B for lipid distribution. The increase of hepatocyte having rneshlike cytoplasm were shown in all hepatic lobules after TX injection and the hepatic plates were disappeared in the region of meshlike hepatocyte aggregation, But the hepatocyte having meshlike cytoplasm were disappeared and hapatic plate were rearranged in CB extract injected mouse. The number of blue black colored lipid drop in hepatic cytoplasm of mouse injected with TX were increased and the size of lipid drop were enlarged. But the number of lipid drop in hepatic cytoplasm of mouse treated CB extract were decreased and the size of lipid drop were diminished. As results indicated that the accumulation of lipid drop caused by TX injection were mitigated by the antihyperlipidermic effect of CB extract.

Supplement of Allicin Prevents the Deterioration of Porcine Oocytes during In Vitro Aging

Yun-Gwi Park,Seung-Eun Lee,Eun-Young Kim,Se-Pill Park 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06

Allicin, one of the chemical substances from garlic, has strong antioxidant activity and considered to represent anti-aging effect in vitro. The objective of this study was to investigate the effects of allicin treatment during porcine oocyte in vitro aging and their in vitro developmental competency after parthenogenetic activation (PA). Porcine oocytes were maturated in vitro for 44 h (control) and 44+24 h with 0, 0.1, 1, 10 and 100 μM allicin (0 AL, 0.1 AL, 1 AL, 10 AL and 100 AL, respectively). We investigated the effects of appropriate concentration of allicin on nuclear, cytoplasmic maturation and the developmental capacity of aged porcine oocytes. Oocyte survival and polar body extrusion were significantly decreased in 0 AL compared with control or 1 AL. The decrease of normal spindle formation, chromosome alignment and expression of maturation marker genes during in vitro aging was prevented in 1 AL. The allicin acted as not anti-oxidant but an autophagy regulator during in vitro aging. After PA, although the cleavage rate were similar among these groups, control, 0 AL and 1 AL, the blastocyst formation was higher in control and 1 AL than 0 AL. Thus, the allicin is effective agent to prevent the deterioration during in vitro aging of porcine oocytes. Therefore, allicin may be helpful for improve the aged oocytes quality to use on in vitro experiments.

β-Cryptoxanthin Supplementation on In Vitro Maturation Medium Strengthens the Quality and Potential of Porcine Oocytes

Yun-Gwi Park,Seung-Eun Lee,Won-Jae Kim,Eun-Young Kim,Se-Pill Park 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & Developmental Biology(Supplement) Vol.41 No.2

One of the reasons about low quality of in vitro matured (IVM) oocytes than those of in vivo is the oxidative stress caused by external oxygen and incomplete antioxidant system. This study was performed to examine the effects of β-cryptoxanthin (BCX, anti- oxidative reagent) on porcine oocyte during IVM and further in vitro developmental potential. The oocytes were matured in IVM medium containing 0, 0.1, 1, 10 and 100 μM BCX (control, 0.1, 1, 10 and 100 B, respectively). The rate of oocyte maturation was higher in 1 B than in control (p<0.1), while that of other BCX treated groups were similar to control. After IVM, the cytoplasmic reactive oxygen species (ROS) expression level in 1 B was the lowest among all groups (p<0.05), while other BCX treated groups were similar to or higher than control. Also, at the classified oocyte maturation stages (GVBD, MⅠ and MⅡ), 1 B significantly decreased ROS and increased GSH level than control (p<0.05). In addition, the relative mRNA expression level of antioxidant genes, SOD1 and PRDX5, were higher in 1 B than in control (p<0.05). As the provitamin A, the relative mRNA expression level of retinoic acid receptor genes, ITGB7 and RXRA, were significantly increased in 1 B than in control (p<0.05). After parthenogenetic activation, the cleavage rates were not different between control and 1 B, however, the blastocyst formation rate was higher in 1 B than in control (p<0.01). In embryo quality, while the DNA fragmentation of blastocysts was similar between control and 1 B, the total cell number (p<0.1) and the relative mRNA expression level of pluripotency marker genes, Pou5f1 and CDX2 (p<0.05), and anti-apoptosis genes, Bcl2L1, Bcl-xl and BIRC5 (p<0.05), were significantly increased in 1 B than in control. These results demonstrate that BCX is helpful for decreasing oxidative stress in porcine oocytes and improves their quality and developmental potential.

β-Cryptoxanthin Treatment during In Vitro Maturation Strengthens the Developmental Potential of Porcine Follicular Oocyte

Yun-Gwi Park,Seung-Eun Lee,Yeo-Jin Son,Min-Young Shin,Sang-Gi Jeong,Eun-Young Kim,Se-Pill Park 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10

Although in vitro production (IVP) techniques of porcine follicular oocytes have progressed and are well studied, the developmental potential of porcine oocytes matured in vitro remains low compared with those matured in vivo. It is well known that one of the reason occurred impair in vitro maturation (IVM) of porcine oocytes is the oxidative stress. Oxidative stress is mainly caused by reactive oxygen species (ROS) generation formed during cellular metabolism. β-cryptoxanthin (BCX) is one of the carotenoid pigment and possesses strong anti-oxidative and free radical scavenging activities and suppresses lipid peroxidation and nitrogen oxide production. The objective of this study was to examine the effects of BCX treatment on porcine oocyte during IVM and their in vitro developmental potential. The follicular oocytes were cultured in IVM medium supplemented with 0, 0.1, 1, 10 and 100 μM BCX (control, 0.1 B, 1 B, 10 B and 100 B). In analysis of intracellular ROS expression level after IVM, 1 B group was the lowest among all groups (p<0.05), while other BCX treated groups are similar to control group. Also, 1 B group was significantly decreased during the classified oocyte maturation stage (GVBD, MⅠ and MⅡ) than control (p<0.05). In addition, the relative mRNA expression level of antioxidant gene (superoxide dismutase-2 and peroxiredoxin-5) was significantly higher in 1 B group than control (p<0.05). After parthenogenetic activation, there was no different in the cleavage rate between two groups, however, the blastocyst formation rate was significantly higher in 1 B group than in control (p<0.05). In embryo quality, the total cell number and DNA fragmentation of blastocysts were no different between two groups. These results demonstrate that BCX is helpful for decreasing ROS level of porcine follicular oocytes and improves their developmental potential.

Phenolics and Antioxidant Properties of 8 Compositae Plants

Gwi Yeong Jang(장귀영),Eun Suk Lee(이은숙),Su Ji Choi(최수지),Hyung Don Kim(김형돈),Min Hye Kang(강민혜),Yun Jeong Ji(지윤정),Yun Ji Lee(이윤지),Geum Soog Kim(김금숙),Seung Eun Lee(이승은) 한국약용작물학회 2022 한국약용작물학회 학술대회논문집 Vol.2022 No.1

β-Cryptoxanthin Treatment during In Vitro Maturation Strengthens the Developmental Potential of Porcine Follicular Oocyte

Yun-Gwi Park,Seung-Eun Lee,Yeo-Jin Son,Min-Young Shin,Sang-Gi Jeong,Eun-Young Kim,Se-Pill Park 한국동물생명공학회(구 한국동물번식학회) 2016 발생공학 국제심포지엄 및 학술대회 Vol.2016 No.10

Effects of Feeder Cell Types on Culture of Mouse Embryonic Stem Cell In Vitro

Yun-Gwi Park,Seung-Eun Lee,Eun-Young Kim,Hyuk Hyun,Min-Young Shin,Yeo-Jin Son,Su-Young Kim,Se-Pill Park 한국발생생물학회 2015 발생과 생식 Vol.19 No.3

The suitable feeder cell layer is important for culture of embryonic stem (ES) cells. In this study, we investigated the effect of two kinds of the feeder cell, MEF cells and STO cells, layer to mouse ES (mES) cell culture for maintenance of stemness. We compare the colony formations, alkaline phosphatase (AP) activities, expression of pluripotency marker genes and proteins of D3 cell colonies cultured on MEF feeder cell layer (D3/MEF) or STO cell layers (D3/STO) compared to feeder free condition (D3/–) as a control group. Although there were no differences to colony formations and AP activities, interestingly, the transcripts level of pluripotency marker genes, Pou5f1 and Nanog were highly expressed in D3/MEF (79 and 93) than D3/STO (61and 77) or D3/– (65 and 81). Also, pluripotency marker proteins, NANOG and SOX-2, were more synthesized in D3/MEF (72.8±7.69 and 81.2±3.56) than D3/STO (32.0±4.30 and 56.0±4.90) or D3/– (55.0±4.64 and 62.0±6.20). These results suggest that MEF feeder cell layer is more suitable to mES cell culture. Key words : Mouse embryonic stem cell, Feeder cell, Pluripotency marker, MEF feeder cell

Electro Cell Fusion Technique as a New Method for Stem Cell Establishment

Yun-Gwi Park,Jun-Beom Lee,Eun-Young Kim,Se-Pill Park 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

Although there are several methods for establishment of stem cell line, most of them has critical limit such as, ethical problem and infectious concern. Accordingly, we investigated the cell fusion technique as a new tool to establish a stem cell line. We cultured mouse embryonic stem cell (ESC) and somatic cells. Then, these two type cells were fused by electro cell fusion that consist of three steps (AC→DC→AC). The fused cells were individually transferred into a 96-well plate and cultured in ESC culture medium for 6 ~ 7 days. Newly formed colonies were evaluated several analysis methods like morphology, alkaline phosphatase (AP) activities, expression of pluripotency marker genes and proteins, and karyotyping. The fusion efficiency from the ESC and somatic cell into colony formation was about 0.3 ~ 0.5 %. The electro cell fused (EF) new stem cell colonies (EF-SC1 ~ 4) were indicated normally round-shape morphology similarly to ESC colonies and each colonies were expressed green fluorescent protein that having somatic cells. Also, all EF-SC groups were highly expressed AP activity and pluripotency marker proteins, POU5f1, NANOG, SOX-2 and SSEA-1. In the transcription levels, all EF-SC groups were significantly higher level of expression in Pou5f1 and Nanog compared to donor cells (ESC and somatic cell) (p<0.05). In particular, the level of Pou5f1 expression was about 2-folds higher in EF-SC2 and EF-SC3 groups than in control and EF-SC1 groups (p<0.05). Also, the level of Nanog expression was very significantly higher in EF-SC2 group (3.5-folds) compared to control ESC group, and the expression levels among treatment groups were variable (ESC<EF-SC1<EF-SC4<EF-SC3<EF-SC2, p<0.05). In karyotype analysis, the results of EF-SC2 and EF-SC3 were presented the same that of ESC, while that of EF-SC1 and EF-SC4 shown aneuploid mutation in chromosome 8. Taken together, these results demonstrate that electro cell fusion technique can be used as a new method to establish of stem cell lines.

In vitro maturation of human pluripotent stem cell-derived cardiomyocyte : A promising approach for cell therapy

Yun-Gwi Park,Yeo-Jin Son,Sung-Hwan Moon,Soon-Jung Park 한국동물생명공학회(구 한국동물번식학회) 2022 Journal of Animal Reproduction and Biotechnology Vol.37 No.2

Currently, there is no treatment to reverse or cure heart failure caused by ischemic heart disease and myocardial infarction despite the remarkable advances in modern medicine. In addition, there is a lack of evidence regarding the existence of stem cells involved in the proliferation and regeneration of cardiomyocytes in adult hearts. As an alternative solution to overcome this problem, protocols for differentiating human pluripotent stem cell (hPSC) into cardiomyocyte have been established, which further led to the development of cell therapy in major leading countries in this field. Recently, clinical studies have confirmed the safety of hPSC-derived cardiac progenitor cells (CPCs). Although several institutions have shown progress in their research on cell therapy using hPSC-derived cardiomyocytes, the functions of cardiomyocytes used for transplantation remain to be those of immature cardiomyocytes, which poses a risk of graft-induced arrhythmias in the early stage of transplantation. Over the last decade, research aimed at achieving maturation of immature cardiomyocytes, showing same characteristics as those of mature cardiomyocytes, has been actively conducted using various approaches at leading research institutes worldwide. However, challenges remain in technological development for effective generation of mature cardiomyocytes with the same properties as those present in the adult hearts. Therefore, in this review, we provide an overview of the technological development status for maturation methods of hPSC-derived cardiomyocytes and present a direction for future development of maturation techniques.

MEF Cell as Feeder Cell Layer Enhances Pluripotency of Mouse Embryonic Stem Cells

Yun-Gwi Park,Seung-Eun Lee,Hyuk Hyun,Min-Young Shin,Yeo-Jin Son,Eun-Young Kim,Se-Pill Park 한국동물생명공학회(구 한국동물번식학회) 2015 발생공학 국제심포지엄 및 학술대회 Vol.2015 No.10