• 제목/요약/키워드: Hepatocyte-derived cell

검색결과 34건 처리시간 0.023초

사람의 제대 및 양막유래 줄기세포의 간세포로의 분화 (Hepatogenic Potential of Umbilical Cord Derived-Stem Cells and Human Amnion Derived-Stem Cells)

  • 김지영;이윤정;박세아;강현미;김경식;조동제;김해권
    • Clinical and Experimental Reproductive Medicine
    • /
    • 제35권4호
    • /
    • pp.247-265
    • /
    • 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.

Human adipose-derived mesenchymal stem cell spheroids improve recovery in a mouse model of elastase-induced emphysema

  • Cho, Ryeon Jin;Kim, You-Sun;Kim, Ji-Young;Oh, Yeon-Mok
    • BMB Reports
    • /
    • 제50권2호
    • /
    • pp.79-84
    • /
    • 2017
  • Emphysema, a pathologic component of the chronic obstructive pulmonary disease, causes irreversible destruction of lung. Many researchers have reported that mesenchymal stem cells can regenerate lung tissue after emphysema. We evaluated if spheroid human adipose-derived mesenchymal stem cells (ASCs) showed greater regenerative effects than dissociated ASCs in mice with elastase-induced emphysema. ASCs were administered via an intrapleural route. Mice injected with spheroid ASCs showed improved regeneration of lung tissues, increased expression of growth factors such as fibroblast growth factor-2 (FGF2) and hepatocyte growth factor (HGF), and a reduction in proteases with an induction of protease inhibitors when compared with mice injected with dissociated ASCs. Our findings indicate that spheroid ASCs show better regeneration of lung tissues than dissociated ACSs in mice with elastase-induced emphysema.

Long-Duration Three-Dimensional Spheroid Culture Promotes Angiogenic Activities of Adipose-Derived Mesenchymal Stem Cells

  • Lee, Jun Hee;Han, Yong-Seok;Lee, Sang Hun
    • Biomolecules & Therapeutics
    • /
    • 제24권3호
    • /
    • pp.260-267
    • /
    • 2016
  • Mesenchymal stem cells (MSCs) offer significant therapeutic promise for various regenerative therapies. However, MSC-based therapy for injury exhibits low efficacy due to the pathological environment in target tissues and the differences between in vitro and in vivo conditions. To address this issue, we developed adipose-derived MSC spheroids as a novel delivery method to preserve the stem cell microenvironment. MSC spheroids were generated by suspension culture for 3 days, and their sizes increased in a time-dependent manner. After re-attachment of MSC spheroids to the plastic dish, their adhesion capacity and morphology were not altered. MSC spheroids showed enhanced production of hypoxia-induced angiogenic cytokines such as vascular endothelial growth factor (VEGF), stromal cell derived factor (SDF), and hepatocyte growth factor (HGF). In addition, spheroid culture promoted the preservation of extracellular matrix (ECM) components, such as laminin and fibronectin, in a culture time- and spheroid size-dependent manner. Furthermore, phosphorylation of AKT, a cell survival signal, was significantly higher and the expression of pro-apoptotic molecules, poly (ADP ribose) polymerase-1 (PARP-1) and cleaved caspase-3, was markedly lower in the spheroids than in MSCs in monolayers. In the murine hindlimb ischemia model, transplanted MSC spheroids showed better proliferation than MSCs in monolayer. These findings suggest that MSC spheroids promote MSC bioactivities via secretion of angiogenic cytokines, preservation of ECM components, and regulation of apoptotic signals. Therefore, MSC spheroid-based cell therapy may serve as a simple and effective strategy for regenerative medicine.

Stevia rebaudiana의 항산화 효과 (Anti-oxidant Effect on Stevia rebaudiana)

  • 정은혜;서혜림;김민규;김영우;조일제
    • 동의생리병리학회지
    • /
    • 제27권6호
    • /
    • pp.764-770
    • /
    • 2013
  • Stevia rebaudiana is a traditional herb used as a sweetener in Brazil and Paraguay as well as Korea and China. This study investigated the efficacy of Stevia rebaudiana methanol extract (SRE) to protect cells against the mitochondrial dysfunction and apoptosis in hepatocyte. To determine the effects of SRE on oxidative stress, we used the human derived hepatocyte cell line, HepG2 cell. Treatment of arachidonic acid (AA)+iron in HepG2 cells synergistically amplified cytotoxicity, as indicated by the excess reactive oxygen species (ROS) and mitochondrial permeability transition by fluorescence activated cell sorter (FACS) and immunoblot analysis. Treatment with SRE protected hepatocytes from AA+iron-induced cellular toxicity, as shown by alterations in the protein levels related with cell viability such as procaspase-3. SRE also prevented the mitochondrial dysfunction induced by AA+iron, and showed anti-oxidant effects as inhibition of $H_2O_2$ production and GSH depletion. Moreover, we measured the effects of SRE on AMP-activated protein kinase (AMPK), a key regulator in determining cell survival or death. Acetyl-CoA Carboxylase (ACC), a direct downstream target of AMPK. SRE increased phosphorylation of ACC, and prevented the inhibition of ACC phosphorylation by AA+iron. These results indicated that SRE has the ability to protect cells against AA+iron-induced $H_2O_2$ production and mitochondrial impairment, which may be mediated with AMPK-ACC pathway.

간 실질세포주에서 니호채(泥胡菜) 분획물이 liver X receptor α 의존적 지방 생성 유전자의 발현에 미치는 효과 (The effects of Hemistepta lyrata Bunge (Bunge) fractionated extract on liver X receptor α-dependent lipogenic genes in hepatocyte-derived cells)

  • 김재광;조일제;김은옥;정대화;구세광;김상찬
    • 대한한의학방제학회지
    • /
    • 제28권3호
    • /
    • pp.255-269
    • /
    • 2020
  • Objectives : Hemistepta lyrata Bunge (Bunge) is a wild herb that has been used for managing fever and wound in Korean Traditional Medicine. The present study explored the effects of H. lyrata extract on liver X receptor (LXR) α-dependent lipogenic genes in hepatocyte-derived cells. Methods : After HepG2 cells or Huh7 cells were pre-treated with 1-10 ㎍/mL of H. lyrata extract or its fractionated extract for 0.5 h, the cells were subsequently exposed to LXR ligand for 6-24 h. Cell viability, LXR response element (LXRE)-driven luciferase activity, sterol regulatory element binding protein-response element (SREBP-RE)-driven luciferase activity, SREBP-1c expression, and mRNA levels of LXRα and its-dependent target genes were determined. In addition, LC-MS/MS analysis was conducted to explore major compounds in H. lyrata-chloroform fractionated extract #4 (HL-CF4). Results : Of various H. lyrata extracts tested, chloroform extract and its fractionated extract #4, HL-CF4, significantly decreased T0901317-mediated SREBP-1c expression. In addition, HL-CF4 significantly reduced LXRE atransactivation and LXRα mRNA expression without any cytotoxicity. Moreover, HL-CF4 prevented the SREBP-RE-driven luciferase activity and mRNA levels of fatty acid synthase and stearoyl-CoA desaturase-1 induced by T0901317. Results from LC-MS/MS analysis at positive/negative mode indicated that HL-CF4 contained several compounds showing m/z 197.1176 (C11H17O3), 693.2913/227.1069 (C38H45O12/C15H15O2), 203.1797 (C15H23), 181.1225 (C11H17O2), 591.2957 (C35H43O8), 379.1040 (C18H19O9), 409.1509 (C20H25O9), 309.1348 (C16H21O6), 391.1404 (C20H23O8), and 669.2924/389.1248 (C36H45O12/C20H21O8). Conclusion : Based on its inhibition of the LXRα-dependent signaling pathway, H. lyrata chloroform extract and HL-CF4 have prophylactic potentials for managing non-alcoholic fatty liver.

삼릉(三稜) 추출물의 간세포 보호 및 미토콘드리아 보호 효과 (Mitochondria protection of Sparganii Rhizoma against oxidative stress in heptocytes)

  • 서혜림;이주희;장미희;권영원;조일제;김광중;박숙자;김상찬;김영우;변성희
    • 대한한의학방제학회지
    • /
    • 제23권2호
    • /
    • pp.189-198
    • /
    • 2015
  • Objectives : Sparganii Rhizoma is frequently used in traditional herbal medicine for treatment of blood stasis, amenorrhea and functional dyspepsia and has been reported to exhibit anti-oxidant, anti-proliferation and anti-angiogenesis peoperties. In this study, we investigated the cytoprotective effect and underlying mechanism of Sparganii Rhizoma water extract (SRE) against oxidative stress-induced mitochondrial dysfunction and apoptosis in hepatocyte. Methods : To determine the effects of SRE on oxidative stress, we induced synergistic cytotoxicity by co-treatment of arachidonic acid (AA) and iron in the HepG2 cell, a human derived hepatocyte cell line. Results : Treatment of SRE increased relative cell viability and altered the expression levels of apoptosis-related proteins such as Bcl-xL, Bcl-2 and procaspase-3. And SRE also inhibited the mitochondrial dysfunction and excessive reactive oxygen species production induced by AA+iron. In addition, SRE activated of AMP-activated protein kinase (AMPK), a potential target for cytoprotection, by increasing the phosphorylation of AMPKα at Thr-172. Morever, SRE increased phosphorylation of acetyl-CoA carboxylase, a direct downstream target of AMPK. Conclusion : These results indicated that SRE has the ability to protect against oxidative stress-induced hepatocyte damage, which may be mediated with AMPK pathway.

The Antimicrobial Insect Peptide CopA3 Blocks Ethanol-Induced Liver Inflammation and Liver Cell Injury in Mice

  • Kim, Ho
    • 한국미생물·생명공학회지
    • /
    • 제50권1호
    • /
    • pp.157-163
    • /
    • 2022
  • Alcoholic liver disease (ALD), which encompasses alcoholic steatosis, alcoholic hepatitis, and alcoholic cirrhosis, is a major cause of morbidity and mortality worldwide. Although the economic and health impacts of ALD are clear, few advances have been made in its prevention or treatment. We recently demonstrated that the insect-derived antimicrobial peptide CopA3 exerts anti-apoptotic and anti-inflammatory activities in various cell systems, including neuronal cells and colonic epithelial cells. Here, we tested whether CopA3 inhibits ethanol-induced liver injury in mice. Mice were intraperitoneally injected with ethanol only or ethanol plus CopA3 for 24 h and then liver injury and inflammatory responses were measured. Ethanol enhanced the production of proinflammatory cytokines, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, interferon (IFN)-γ, and IL-10. It also induced hepatocyte apoptosis and ballooning degeneration in hepatocytes. Notably, all these effects were eliminated or significantly reduced by CopA3 treatment. Collectively, our findings demonstrate that CopA3 ameliorates ethanol-induced liver cell damage and inflammation, suggesting the therapeutic potential of CopA3 for treating ethanol-induced liver injury.

Involvement of NOX2-derived ROS in human hepatoma HepG2 cell death induced by Entamoeba histolytica

  • Young Ah Lee ;Myeong Heon Shin
    • Parasites, Hosts and Diseases
    • /
    • 제61권4호
    • /
    • pp.388-396
    • /
    • 2023
  • Entamoeba histolytica is an enteric tissue-invasive protozoan parasite causing amoebic colitis and liver abscesses in humans. Amoebic contact with host cells activates intracellular signaling pathways that lead to host cell death via generation of caspase-3, calpain, Ca2+ elevation, and reactive oxygen species (ROS). We previously reported that various NADPH oxidases (NOXs) are responsible for ROS-dependent death of various host cells induced by amoeba. In the present study, we investigated the specific NOX isoform involved in ROS-dependent death of hepatocytes induced by amoebas. Co-incubation of hepatoma HepG2 cells with live amoebic trophozoites resulted in remarkably increased DNA fragmentation compared to cells incubated with medium alone. HepG2 cells that adhered to amoebic trophozoites showed strong dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, suggesting intracellular ROS accumulation within host cells stimulated by amoebic trophozoites. Pretreatment of HepG2 cells with the general NOX inhibitor DPI or NOX2-specific inhibitor GSK 2795039 reduced Entamoeba-induced ROS generation. Similarly, Entamoeba-induced LDH release from HepG2 cells was effectively inhibited by pretreatment with DPI or GSK 2795039. In NOX2-silenced HepG2 cells, Entamoeba-induced LDH release was also significantly inhibited compared with controls. Taken together, the results support an important role of NOX2-derived ROS in hepatocyte death induced by E. histolytica.

VEGF와 HGF에 의해 유도된 U-373-MG 세포의 이동 및 침윤에 미치는 단백질분해효소의 효과 (Effect of Proteases on the Migration and Invasion of U-373-MG Cells Induced by Vascular Endothelial Growth Factor and Hepatocyte Growth Factor)

  • 전희영;김환규
    • 생명과학회지
    • /
    • 제26권10호
    • /
    • pp.1189-1195
    • /
    • 2016
  • Vascular endothelial growth factor (VEGF)와 hepatocyte growth factor (HGF)는 강력한 혈관신생 유도인자로 알려져 있다. 세포가 이동하고 침윤하기 위해서는 세포의 증식과 더불어 주변의 세포외기질을 분해하는 단백질분해효소의 분비가 선행되어야 한다. 본 연구에서는 인간의 악성신경교종 유래 세포주인 U-373-MG 세포에 VEGF와 HGF를 처리하여 세포의 증식, matrix metalloproteinase-2 (MMP-2)와 MMP-9 및 플라스민의 분비, 세포의 이동 및 침윤에 미치는 효과를 조사하였다. 또한 단백질분해효소 억제제 처리를 통하여 세포의 증식, 이동 및 침윤에 미치는 단백질분해효소의 역할을 조사하였다. 연구 결과, VEGF와 HGF의 병용처리 시 VEGF와 HGF의 단독 처리 시보다 세포의 증식, MMP-2, MMP-9 및 플라스민의 분비, 세포의 이동 및 침윤이 유의할만하게 증가되었다. 한편 VEGF와 HGF 처리에 의한 U-373-MG 세포의 증식, 이동 및 침윤 증가에 미치는 단백질분해효소의 효과를 MMPs 억제제인 BB-94를 처리하여 조사한 결과 최대 이동 효과를 나타낸 HGF와 VEGF의 병용처리군 보다 세포의 이동이 32% 억제되었고 플라스민 억제제인 α2AP에 의해서도 29% 억제되었다. 또한 U-373-MG 세포의 침윤 역시 BB-94와 α2AP 처리에 의해 유의할 만하게 억제되었다. 이러한 결과는 VEGF와 HGF에 의한 MMP-2, MMP-9 및 플라스민의 분비증가에 의해 직접 또는 간접적인 경로를 통하여 U-373-MG 세포의 증식, 이동 및 침윤을 증가시킨다고 여겨진다.

Evaluation of 1-DNJ extracted from Silkworm in GBV-B Infected Marmoset Hepatocytes

  • James R. Jacob;Yoo, Jung-Eun;Seong, Su-Il;Ryu, Kang-Sun;Kim, Young-Ho
    • 한국잠사학회:학술대회논문집
    • /
    • 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
    • /
    • pp.53-53
    • /
    • 2003
  • Alpha-glucosidase inhibitor, 1-deoxynojirimycin, has been shown to eliminate the production of ER-budding viruses in the ER of host cell. This glucose-derived iminosugar derivative, 1-DNJ, has been largely purified from midgut of silkworm, Bombyx mori, Here, we have screened the antiviral activity in the GBV-B infected marmoset hepatocyte cell line. (omitted)

  • PDF