• 한국발생생물학회지:발생과생식
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• 제12권2호
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• pp.183-194
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• 2008

A variety of stem cells has been emerging as therapeutic cells that can replace organ transplantation in human liver diseases. The present study focused on whether human eyelid adipose-derived stem cells (HAD) might differentiate into functional hepatocyte-like cells in vitro. HAD were isolated from human eyelid adipose tissue. Effect of dimethyl sulfoxide (DMSO), fibroblast growth factor (FGF)-2 and FGF-4 on the hepatic differentiation of HAD have been examined in vitro. Immunocytochemical analysis and PAS staining showed that HAD cultured in both DMSO and FGF-4 exhibited the most intense staining than HAD of the other experimental groups. These HAD expressed numerous hepatocyte-related genes. Immunoblotting analyses showed that HAD cultured in the presence of DMSO and FGF-4 secreted higher amount of human albumin than HAD cultured in other conditions. Urea analysis also demonstrated that these HAD produced higher amount of urea than any other groups of HAD. In conclusion, combined treatment of DMSO and FGF-4 could effectively induce the functional differentiation of HAD into hepatocyte-like cells.

• Molecules and Cells
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• 제41권7호
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• pp.639-645
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• 2018

Liver transplantation is recommended for patients with liver failure, but liver donors are limited. This necessitates the development of artificial livers, and hepatocytes are necessary to develop such artificial livers. Although induced hepatocyte-like cells are used in artificial livers, the characteristics of mouse induced hepatocyte-like cells (miHeps) reprogrammed with embryonic fibroblasts have not yet been clarified. Therefore, this study investigated the mechanisms underlying the survival, function, and death of miHeps. miHeps showed decreased cell viability, increased cytotoxicity, decreased hepatic function, and albumin and urea secretion at passage 14. Addition of necrostatin-1 (NEC-1) to miHeps inhibited necrosome formation and reactive oxygen species generation and increased cell survival. However, NEC-1 did not affect the hepatic function of miHeps. These results provide a basis for development of artificial livers using hepatocytes.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
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• pp.76-77
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• 2006

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2006년도 The 5th Biannual Meeting of Pacific Rim Society for Fertility and Sterility
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• pp.172.1-172.1
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• 2006

• Clinical and Experimental Reproductive Medicine
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• 제35권4호
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• pp.247-265
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• 2008

목 적: 사람의 제대유래 줄기세포 (HUC)와 양막유래 줄기세포 (HAM)를 간유사세포로 분화 시키고자 하였다. 연구방법: 산모의 동의아래 만삭 정상 산모로부터 양막 및 제대를 얻고 줄기세포를 분리, 배양하였다. 분리된 세포의 줄기세포로서의 특성을 규명하기 위하여 RT-PCR, 세포면역화학 분석 그리고 중배엽성세포로의 분화능 실험을 하였다. 또한 이들 줄기세포를 분화 배양액에서 간유사세포로 분화 유도 후, 간세포 특이 유전자 발현, 알부민 ELISA, 알부민에 대한 면역블로팅 및 세포면역화학염색 그리고 PAS 염색을 시행하였다. 결 과: 사람의 양막 및 제대로부터 세포를 분리하여 줄기세포로서의 특성을 조사하고, 이들 세포의 지방세포, 연골세포, 골아세포로의 분화능력을 확인하였다. 동일한 분화 배양액에서 HUC과 HAM을 간유사세포로 분화시킨 결과 HUC이 모든 조건에서 HAM보다 알부민과 요소를 더 많이 합성 분비하였다. 간유사세포로의 분화능력이 더 좋은 것으로 나타난 HUC을 가지고 최적의 분화를 유도한 결과 $1.2{\pm}0.8\;{\mu}g/mL$의 알부민을 합성 분비하였고, $8.9{\pm}1.2\;mg/dL$의 요소를 합성 분비하였다. 결 론: HUC과 HAM 모두 알부민을 분비하는 간유사세포로 분화할 수 있었으며, HUC이 더 잘 유도되는 것으로 관찰되었다. 이러한 결과로써 HUC과 HAM을 간질환 환자의 간세포 이식 치료에 필요한 세포치료제로써 이용할 수 있을 것으로 생각된다.

• BMB Reports
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• 제55권12호
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• pp.633-638
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• 2022

Liver regeneration is a well-known systemic homeostatic phenomenon. The N⁶-methyladenosine (m⁶A) modification pathway has been associated with liver regeneration and hepatocellular carcinoma. m⁶A methyltransferases, such as methyltransferase 3 (METTL3) and methyltransferase 14 (METTL14), are involved in the hepatocyte-specific-regenerative pathway. To illustrate the role of METTL14, secreted from non-parenchymal liver cells, in the initiation phase of liver regeneration, we performed 70% partial hepatectomy (PH) in Mettl14 heterozygous (HET) and wild-type (WT) mice. Next, we analyzed the ratio of liver weight to body weight and the expression of mitogenic stimulators derived from non-parenchymal liver cells. Furthermore, we evaluated the expression of cell cycle-related genes and the hepatocyte proliferation rate via MKI67-immunostaining. During regeneration after PH, the weight ratio was lower in Mettl14 HET mice compared to WT mice. The expressions of hepatocyte growth factor (HGF) and tumor necrosis factor (TNF)-α, mitogens derived from non-parenchymal liver cells that stimulate the cell cycle, as well as the expressions of cyclin B1 and D1, which regulate the cell cycle, and the number of MKI67-positive cells, which indicate proliferative hepatocyte in the late G1-M phase, were significantly reduced in Mettl14 HET mice 72 h after PH. Our findings demonstrate that global Mettl14 mutation may interrupt the homeostasis of liver regeneration after an acute injury like PH by restraining certain mitogens, such as HGF and TNF-α, derived from sinusoidal endothelial cells, stellate cells, and Kupffer cells. These results provide new insights into the role of METTL14 in the clinical treatment strategies of liver disease.

• Bulletin of the Korean Chemical Society
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• 제33권6호
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• pp.1983-1988
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• 2012

Stem cell transplantation is emerging as a possible new treatment for liver cirrhosis, and recent animal studies have documented the benefits of stem cell therapy in a hepatic fibrosis model. However, the underlying mechanism of stem cell therapy is still unclear. Among the proposed mechanisms, the cell replacement mechanism is the oldest and most important, in which permanently damaged tissue can be replaced by normal tissue to restore function. In the present study, Cy5.5-labeled superparamagnetic iron oxide (SPIO) was used to label human mesenchymal stem cells. The uptake of fluorescently labeled nanoparticles enabled the detection and monitoring of the transplanted stem cells; therefore, we confirmed the direct incorporation and differentiation of SPIO into the hepatocyte-like transplanted stem cells by detecting human tyrosine aminotransferase (TAT), well-known enzymatic marker for hepatocyte-specific differentiation.

• Archives of Plastic Surgery
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• 제39권6호
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• pp.593-599
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• 2012

Background This study aimed to investigate the possibility of isolating mesenchymal stem cells (MSCs) from human thigh adipose tissue and the ability of human thigh adipose stem cells (HTASCs) to differentiate into hepatocytes. Methods The adipose-derived stem cells (ADSCs) were isolated from thigh adipose tissue. Growth factors, cytokines, and hormones were added to the collagen coated dishes to induce the undifferentiated HTASCs to differentiate into hepatocyte-like cells. To confirm the experimental results, the expression of hepatocyte-specific markers on undifferentiated and differentiated HTASCs was analyzed using reverse transcription polymerase chain reaction and immunocytochemical staining. Differentiation efficiency was evaluated using functional tests such as periodic acid schiff (PAS) staining and detection of the albumin secretion level using enzyme-linked immunosorbent assay (ELISA). Results The majority of the undifferentiated HTASCs were changed into a more polygonal shape showing tight interactions between the cells. The differentiated HTASCs up-regulated mRNA of hepatocyte markers. Immunocytochemical analysis showed that they were intensely stained with anti-albumin antibody compared with undifferentiated HTASCs. PAS staining showed that HTASCs submitted to the hepatocyte differentiation protocol were able to more specifically store glycogen than undifferentiated HTASCs, displaying a purple color in the cytoplasm of the differentiated HTASCs. ELISA analyses showed that differentiated HTASCs could secrete albumin, which is one of the hepatocyte markers. Conclusions MSCs were islolated from human thigh adipose tissue differentiate to heapatocytes. The source of ADSCs is not only abundant abdominal adipose tissue, but also thigh adipose tissue for cell therapy in liver regeneration and tissue regeneration.

• 한국발생생물학회지:발생과생식
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• 제10권1호
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• pp.63-73
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• 2006

간질환 환자의 대부분은 간 조직 손상으로 인해 간세포의 재생 능력이 감소한다. 간세포 이식은 이러한 간질환을 치료하는데 있어 혁신적인 방법으로 대두되고 있으나, 여전히 많은 의문과 문제점이 제기되고 있다. 사람의 양막으로부터 얻은 줄기 세포를 이용하여 간세포 분화를 위한 최적의 조건을 알아 보고자 하였다. 세포내 알부민에 대한 면역 화학적 방법, 세포내 글리코겐의 특이 염색법, 세포의 형태적 변화 연구 방법 등을 이용하여 여러가지 배양 조건을 조사한 결과, 배양 접시를 fibronectin으로 coating하고 배양액내에 insulin/transferrin/selenium(ITS)을 첨가하는 것이 양막 줄기세포의 간세포로의 분화에 효과적이었다. 또한 배양액내에 fibroblast growth factor(FGF)-1과 FGF-2를 함께 첨가하는 것이 둘 중 하나만 첨가하거나 첨가하지 않는 것보다 효과적이었다. 한편 분화 배양은 한가지 배양액을 사용한 지속적인 배양법(continuous culture method)보다 배양 조건을 달리하여 두 단계로 배양하는 2단계 배양법(two-step culture method)가 훨씬 효과적이었다. 마지막으로, 기본 배양액에 FGF-2와 FGF-4를 첨가한 조건과 FGF-4와 $TGF-{\alpha}$를 첨가한 조건이 다른 조건 보다 알부민 분비를 많이 하는 것으로 보아 FGF-4가 간세포 분화 과정에 중요한 역할을 하는 것으로 여겨지며 FGF-2 및 $TGF-{\alpha}$ 첨가는 더욱 효과적인 배양 조건으로 관찰되었다. 따라서, 양막에서 유래한 성체 줄기 세포는 적절한 배양 조건이 주어질 때, 간세포로 분화가 가능하며, 분화 과정에서 FGF-4가 주도적인 역할을 하며 FGF-2와 $TGF-{\alpha}$는 상승 효과를 갖는 것으로 사료된다.

• 대한생식의학회:학술대회논문집
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• 대한생식의학회 2007년도 제53차 추계학술대회 초록집
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• pp.112.1-112.1
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• 2007