• 제목/요약/키워드: in vitro Differentiation

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In Vitro Neural Cell Differentiation Derived from Human Embryonic Stem Cells: II. Generation of Specific Neurons from Neural Progenitor Cells Treated with BDNF and PDGF

  • Jo Hyeon-Jeong;Kim Eun-Yeong;Choe Gyeong-Hui;An So-Yeon;Park Se-Pil;Im Jin-Ho
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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    • pp.84-84
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    • 2002
  • This study was to investigate generation of the specific neuronal cell in vitro from the neural progenitors derived from human embryonic stem (hES, MB03) cells. For the neural progenitor cell formation, we produced embryoid bodies (EB: for 5 days, without mitogen) from hES cells and then neurospheres (for 7-10 days, 20 ng/㎖ of bFGF added N2 medium) from EB. And then for the differentiation into neuronal cells, neural progenitor cells were cultured in N2 medium (without bFGF) supplemented with brain derived neurotrophic factor (BDNF, 5 ng/㎖) or platelet derived growth factor-bb (pDGF-bb, 20ng/㎖) for 2 weeks. (omitted)

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Anti-adipogenic Activity of Acer tegmentosum and its Constituent, Catechin in 3T3-L1 Cells

  • Liu, Qing;Shin, Eun-Jin;Ahn, Mi-Jeong;Hwang, Bang-Yeon;Lee, Mi-Kyeong
    • Natural Product Sciences
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    • 제17권3호
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    • pp.212-215
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    • 2011
  • In the course of screening anti-adipogenic activity of natural products employing the preadipocyte cell line, 3T3-L1 as an in vitro assay system, the EtOAc fraction of the stem barks of Acer tegmentosum Maxim (Aceraceae) showed significant inhibitory activity on adipocyte differentiation as assessed by measuring fat accumulation using Oil Red O staining. Activity-guided fractionation led to the isolation of active constituent, (+)-catechin. (+)-Catechin showed inhibitory activity on adipocyte differentiation in dose-dependent manner. Further studies with interval treatment demonstrated that (+)-catechin exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Our present study also showed that (+)-catechin significantly inhibited the preadipocyte proliferation. Taken together, these results suggest that (+)-catechin, a constituent of A. tegmentosum might contribute the anti-adipogenic activity of A. tegmentosum.

Metformin enhances the osteogenic activity of rat bone marrow mesenchymal stem cells by inhibiting oxidative stress induced by diabetes mellitus: an in vitro and in vivo study

  • Kai Dong;Wen-Juan Zhou;Zhong-Hao Liu
    • Journal of Periodontal and Implant Science
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    • 제53권1호
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    • pp.54-68
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    • 2023
  • Purpose: The purpose of this study was to determine whether metformin (MF) could alleviate the expresssion of reactive oxygen species (ROS) and improve the osteogenic ability of bone marrow mesenchymal stem cells derived from diabetic rats (drBMSCs) in vitro, and to evaluate the effect of MF on the ectopic osteogenesis of drBMSCs in a nude mouse model in vivo. Methods: BMSCs were extracted from normal and diabetic rats. In vitro, a cell viability assay (Cell Counting Kit-8), tests of alkaline phosphatase (ALP) activity, and western blot analysis were first used to determine the cell proliferation and osteogenic differentiation of drBMSCs that were subjected to treatment with different concentrations of MF (0, 50, 100, 200, 500 µM). The cells were then divided into 5 groups: (1) normal rat BMSCs (the BMSCs derived from normal rats group), (2) the drBMSCs group, (3) the drBMSCs + Mito-TEMPO (10 µM, ROS scavenger) group, (4) the drBMSCs + MF (200 µM) group, and (5) the drBMSCs + MF (200 µM) + H2O2 (50 µM, ROS activator) group. Intracellular ROS detection, a senescence-associated β-galactosidase assay, ALP staining, alizarin red staining, western blotting, and immunofluorescence assays were performed to determine the effects of MF on oxidative stress and osteogenic differentiation in drBMSCs. In vivo, the effect of MF on the ectopic osteogenesis of drBMSCs was evaluated in a nude mouse model. Results: MF effectively reduced ROS levels in drBMSCs. The cell proliferation, ALP activity, mineral deposition, and osteogenic-related protein expression of drBMSCs were demonstrably higher in the MF-treated group than in the non-MF-treated group. H2O2 inhibited the effects of MF. In addition, ectopic osteogenesis was significantly increased in drBMSCs treated with MF. Conclusions: MF promoted the proliferation and osteogenic differentiation of drBMSCs by inhibiting the oxidative stress induced by diabetes and enhenced the ectopic bone formation of drBMSCs in nude mice.

Acteoside induce antiproliferation and differentiation on HL-60, Human leukemia cell line, by cell cycle arrest.

  • Lee, Kyoung-Won;Choi, Jung-Hye;Lee, Kyung-Tae;Lee, Yong-Sup;Kim, Hyoung-Ja;Park, Hee-Juhn
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.215.1-215.1
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    • 2003
  • We investigated the in vitro effect of Acteoside , phenylpropanoid glycosides. is a natural product isolated from …. on proliferation, differentiation and cell cycle regulation in human promyelocytic HL -60 leukemia cells. Acteoside significantly inhibited the proliferation of HL -60 cells, with IC50 of about 30$\mu\textrm{g}$/$m\ell$. It was also found to be a potent inducer of differentiation in human leukemia derived HL-60 cells through the examination of differentiation markers. (omitted)

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랫드 전배아배양법을 이용한 2-Bromopropane의 최기형성 평가 (Teratogenicity Evaluation of 2-Bromopropane Using Rat Whole Embryo Culture)

  • 김종춘;신동호;김성호;양영수;오기석;강성철;정문구
    • Toxicological Research
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    • 제22권2호
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    • pp.127-133
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    • 2006
  • Recently, we have reported that the environmental pollutant 2-bromopropane (2-BP) induces a significant embryo-fetal developmental toxicity in rats. However, the cause of developmental toxicity and the relationship between maternal and developmental toxicities could not be elucidated because the developmental toxicity of 2-BP was observed only in the presence of maternal toxicity The in vitro teratogenicity study using whole embryo culture was carried out to understand the teratogenic properties and the possible mechanism of teratogenicity induced by 2-BP in rats. Rat embryos aged 9.5 days were cultured in vitro for 48 hrs at medium concentrations of 0, 1, 3, or 10 mg/ml of 2-BP. Embryos were evaluated for growth, differentiation, and morphological alterations at the end of the culture period. At 10 mg/ml, 2-BP caused a delay in the growth and differentiation of embryos and an increase in the incidence of morphological alterations, including altered yolk sac circulation, abnormal axial rotation, craniofacial hypoplasia, open neuropore, absent optic vesicle and kinked somites. At 3 mg/ml, only a delay in the growth and differentiation of embryos was observed. There were no adverse effects on embryonic growth and development at the concentration of 1 mg/ml. The results showed that the exposure of 2-BP to rat embryos results in a developmental delay and morphological alterations at dose levels of 3 mg/ml culture media or higher and that 2-BP can induce a direct developmental toxicity in rat embryos.

Inhibitory Activity of Three Varieties of Adlay (Coix Seed) on Adipocyte Differentiation in 3T3-L1 cells

  • Lee, Mi-Kyeong;Shin, Eun-Jin;Liu, Qing;Hwang, Bang-Yeon;Lee, Jung-Bok;Kim, Sun-Yeou;Lee, Jae-Hak
    • Natural Product Sciences
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    • 제16권4호
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    • pp.291-294
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    • 2010
  • In obesity, adipocytes undergo abnormal growth characterized by increased cell numbers and differentiation. Thus, inhibition of mitogenesis of preadipocytes and their differentiation to adipocytes would be beneficial for the prevention and progression of obesity. In the present study, we attempted to evaluate antiadipogenic activity of adlay (Coix seed, the seed of Coix lacryma-jobi L. var. ma-yuen Stapf) employing preadipocytes cell line, 3T3-L1 as an in vitro assay system. Because several varieties of adlay are in use in Korea, anti-adipogenic activity of three varieties of adlay such as Sang-Gang, Jo-Hyun and Yulmu-Ilho was evaluated. All the three varieties of adlay showed significant inhibitory activity on adipocyte differentiation as assessed by measuring fat accumulation using Oil Red O staining. Adlay, however, showed little effects on adipocyte proliferation. Further studies with interval treatment demonstrated that adlay exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Taken together, adlay might be useful in the prevention of obesity.

Osteoclast Differentiation Factor Engages the PI 3-kinase, p38, and ERK pathways for Avian Osteoclast Differentiation

  • Kim, Hong-Hee;Kim, Hyun-Man;Kwack, Kyu-Bum;Kim, Si-Wouk;Lee, Zang-Hee
    • BMB Reports
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    • 제34권5호
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    • pp.421-427
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    • 2001
  • Osteoclasts, cells primarily involved in bone resorption, originate from the hematopoietic precursor cells of the monocyte/macrophage lineage and differentiate into multinucleated mature forms. We developed an in vitro osteoclast culture system using embryonic chicken bone marrow cells. This culture system can be utilized in studies on the differentiation and function of osteoclasts. Phosphatidylinositol 3-kinase (PI3-kinase) and mitogen-activated protein kinases (MAPKs) have been implicated in diverse cellular functions including proliferation, migration, and survival. Using the developed avian osteoclast culture system, we examined the involvement of these kinases in osteoclast differentiation by employing specific inhibitors of the kinases. We Found that the inhibition of the PI 3-kinase, p38, or ERK interfered with osteoclast formation, suggesting that the signaling pathways that involve these molecules participate in the process of chicken osteoclast differentiation.

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Efficient In Vitro Labeling Rabbit Bone Marrow-Derived Mesenchymal Stem Cells with SPIO and Differentiating into Neural-Like Cells

  • Zhang, Ruiping;Li, Jing;Li, Jianding;Xie, Jun
    • Molecules and Cells
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    • 제37권9호
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    • pp.650-655
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    • 2014
  • Mesenchymal stem cells (MSCs) can differentiate into neural cells to treat nervous system diseases. Magnetic resonance is an ideal means for cell tracking through labeling cells with superparamagnetic iron oxide (SPIO). However, no studies have described the neural differentiation ability of SPIO-labeled MSCs, which is the foundation for cell therapy and cell tracking in vivo. Our results showed that bone marrow-derived mesenchymal stem cells (BM-MSCs) labeled in vitro with SPIO can be induced into neural-like cells without affecting the viability and labeling efficiency. The cellular uptake of SPIO was maintained after labeled BM-MSCs differentiated into neural-like cells, which were the basis for transplanted cells that can be dynamically and non-invasively tracked in vivo by MRI. Moreover, the SPIO-labeled induced neural-like cells showed neural cell morphology and expressed related markers such as NSE, MAP-2. Furthermore, whole-cell patch clamp recording demonstrated that these neural-like cells exhibited electrophysiological properties of neurons. More importantly, there was no significant difference in the cellular viability and $[Ca^{2+}]_i$ between the induced labeled and unlabeled neural-like cells. In this study, we show for the first time that SPIO-labeled MSCs retained their differentiation capacity and could differentiate into neural-like cells with high cell viability and a good cellular state in vitro.