• Title/Summary/Keyword: Tissue differentiation

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Differentiation of Human Adult Adipose Derived Stem Cell in vitro and Immunohistochemical Study of Adipose Derived Stem Cell after Intracerebral Transplantation in Rats

  • Ko, Kwang-Seok;Lee, Il-Woo;Joo, Won-Il;Lee, Kyung-Jun;Park, Hae-Kwan;Rha, Hyung-Keun
    • Journal of Korean Neurosurgical Society
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    • v.42 no.2
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    • pp.118-124
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    • 2007
  • Objective : Adipose tissue is derived from the embryonic mesoderm and contains a heterogenous stromal cell population. Authors have tried to verify the characteristics of stem cell of adipose derived stromal cells (ADSCs) and to investigate immunohistochemical findings after transplantation of ADSC into rat brain to evaluate survival, migration and differentiation of transplanted stromal cells. Methods : First ADSCs were isolated from human adipose tissue and induced adipose, osseous and neuronal differentiation under appropriate culture condition in vitro and examined phenotypes profile of human ADSCs in undifferentiated states using flow cytometry and immunohistochemical study. Human ADSCs were transplanted into the healthy rat brain to investigate survival, migration and differentiation after 4 weeks. Results : From human adipose tissue, adipose stem cells were harvested and subcultured for several times. The cultured ADSCs were differentiated into adipocytes, osteoctye and neuron-like cell under conditioned media. Flow cytometric analysis of undifferentiated ADSCs revealed that ADSCs were positive for CD29, CD44 and negative for CD34, CD45, CD117 and HLA-DR. Transplanted human ADSCs were found mainly in cortex adjacent to injection site and migrated from injection site at a distance of at least 1 mm along the cortex and corpus callosum. A few transplanted cells have differentiated into neuron and astrocyte. Conclusion : ADSCs were differentiated into multilineage cell lines through transdifferentiation. ADSCs were survived and migrated in xenograft without immunosuppression. Based on this data, ADSCs may be potential source of stem cells for many human disease including neurologic disorder.

Propyl Gallate Inhibits Adipogenesis by Stimulating Extracellular Signal-Related Kinases in Human Adipose Tissue-Derived Mesenchymal Stem Cells

  • Lee, Jeung-Eun;Kim, Jung-Min;Jang, Hyun-Jun;Lim, Se-Young;Choi, Seon-Jeong;Lee, Nan-Hee;Suh, Pann-Ghill;Choi, Ung-Kyu
    • Molecules and Cells
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    • v.38 no.4
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    • pp.336-342
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    • 2015
  • Propyl gallate (PG) used as an additive in various foods has antioxidant and anti-inflammatory effects. Although the functional roles of PG in various cell types are well characterized, it is unknown whether PG has effect on stem cell differentiation. In this study, we demonstrated that PG could inhibit adipogenic differentiation in human adipose tissue-derived mesenchymal stem cells (hAMSCs) by decreasing the accumulation of intracellular lipid droplets. In addition, PG significantly reduced the expression of adipocyte-specific markers including peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$), CCAAT enhancer binding protein-${\alpha}$ (C/EBP-${\alpha}$), lipoprotein lipase (LPL), and adipocyte fatty acid-binding protein 2 (aP2). PG inhibited adipogenesis in hAMSCs through extracellular regulated kinase (ERK) pathway. Decreased adipogenesis following PG treatment was recovered in response to ERK blocking. Taken together, these results suggest a novel effect of PG on adipocyte differentiation in hAMSCs, supporting a negative role of ERK1/2 pathway in adipogenic differentiation.

Tumor Necrosis factor-α Promotes Osteogenesis of Human Bone Marrow-derived Mesenchymal Stem Cells through JNK-dependent Pathway (Tumor necrosis factor-α에 의한 골수 유래 중간엽 줄기세포의 골세포로의 분화 촉진에서 JNK의 역할)

  • Kim, Mi-Ra;Song, Hae-Young;Kim, Jae-Ho
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1207-1213
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    • 2006
  • Tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ has been implicated in skeletal diseases by promoting bone loss in inflammatory bone diseases. In the present study, we examined the effects of $TNF-{\alpha}$ on osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs). $TNF-{\alpha}$ dose-dependently promoted matrix mineralization of hBMSCs with a maximal stimulation at 2ng/ml. $TNF-{\alpha}$ increased expression of alkaline phosphatase, which plays a crucial role for the matrix deposition. The $TNF-{\alpha}-stimulated$ osteoblastic differentiation was not affected by $NF_kB$ inhibitors, BAY and SN50. However, a JNK-specific inhibitor, SP600125 completely abolished the $TNF-{\alpha}-stimulated$ matrix mineralization and expression of alkaline phosphatase. These results suggest that $TNF-{\alpha}$ enhances osteoblastic differentiation of hBMSCs through JNK-dependent pathway.

High Frequency Regeneration of Plantlets from Seedling Explants of Asteracantha longifolia (L.) NEES

  • Mishra Ramya Ranjan;Behera Motilal;Kumar Deep Ratan;Panigrahi Jogeswar
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.27-35
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    • 2006
  • Plantlet regeneration in Asteracantha longifolia(L.) Nees (Acanthaceae), a medicinal herb has been achieved from seedling explants on basal MS medium. Three different seedling explants including node, internode and leaf segments on used. Of these three explant, leaf explants gave better response for both callus mediated organogenesis and direct multiple shoot induction. Number of explants showing differentiation of shout buds was higher on MS media supplemented with BA compared to kinetin. MS medium fortified with BA ($2.0mgl^{-1}$) and NAA ($0.5mgl^{-1}$) was found to be most suitable for both callus mediated organogenesis and elongation of shouts. The elongated shoots were successfully routed on MS medium fortified with NAA or IBA. Among them $0.1mgl^{-1}$ NAA or $0.2mgl^{-1}$ IBA provides better response for rhizogenesis. Regenerated plantlets were successfully established in soil where 85.4% or them developed into morphologically normal and fertile plants. RAPD profiling using four decamer primers confirmed the genetic uniformity of the regenerated plantlets and substantiated the efficacy and suitability of this protocol for in vitro propagation of A. longifolia.

Tracking of Stem Cells from Human Exfoliated Deciduous Teeth Labeled with Molday ION Rhodamine-B during Periodontal Bone Regeneration in Rats

  • Nan Zhang;Li Xu;Hao Song;Chunqing Bu;Jie Kang;Chuanchen Zhang;Xiaofei Yang;Fabin Han
    • International Journal of Stem Cells
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    • v.16 no.1
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    • pp.93-107
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    • 2023
  • Background and Objectives: Chronic periodontitis can lead to alveolar bone resorption and eventually tooth loss. Stem cells from exfoliated deciduous teeth (SHED) are appropriate bone regeneration seed cells. To track the survival, migration, and differentiation of the transplanted SHED, we used super paramagnetic iron oxide particles (SPIO) Molday ION Rhodamine-B (MIRB) to label and monitor the transplanted cells while repairing periodontal bone defects. Methods and Results: We determined an appropriate dose of MIRB for labeling SHED by examining the growth and osteogenic differentiation of labeled SHED. Finally, SHED was labeled with 25 ㎍ Fe/ml MIRB before being transplanted into rats. Magnetic resonance imaging was used to track SHED survival and migration in vivo due to a low-intensity signal artifact caused by MIRB. HE and immunohistochemical analyses revealed that both MIRB-labeled and unlabeled SHED could promote periodontal bone regeneration. The colocalization of hNUC and MIRB demonstrated that SHED transplanted into rats could survive in vivo. Furthermore, some MIRB-positive cells expressed the osteoblast and osteocyte markers OCN and DMP1, respectively. Enzyme-linked immunosorbent assay revealed that SHED could secrete protein factors, such as IGF-1, OCN, ALP, IL-4, VEGF, and bFGF, which promote bone regeneration. Immunofluorescence staining revealed that the transplanted SHED was surrounded by a large number of host-derived Runx2- and Col II-positive cells that played important roles in the bone healing process. Conclusions: SHED could promote periodontal bone regeneration in rats, and the survival of SHED could be tracked in vivo by labeling them with MIRB. SHED are likely to promote bone healing through both direct differentiation and paracrine mechanisms.

Role of NFAT5 in Osteogenic Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells (인체 지방 유래 중간엽 줄기세포의 골분화 조절 기전에서 NFAT5의 역할)

  • Lee, Sun Young;Yang, Ji won;Jung, Jin Sup
    • Journal of Life Science
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    • v.23 no.4
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    • pp.471-478
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    • 2013
  • Human adipose tissue-derived mesenchymal stem cells (hADSCs) have therapeutic potential, including the ability to self-renew and differentiate into multiple lineages. Understanding of molecular mechanisms of stem cell differentiation is important for improving the therapeutic efficacies of stem cell transplantation. In this study, we determined the role of nuclear factor of activated T cells (NFAT5) in the osteogenic differentiation of hADSCs. The down-regulation of NFAT5 expression by the transfection of a specific siRNA significantly inhibited osteogenic differentiation of hADSCs and decreased the activity of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$) promoter without affecting their proliferation and adipogenic differentiation. The inhibition of NFAT5 expression inhibited the basal and Tumor Necrosis Factor ${\alpha}$ (TNF-${\alpha}$) induced activation of NF-${\kappa}B$, but it did not affect TNF-${\alpha}$-induced degradation of the $I{\kappa}B$ protein. These findings indicate that NFAT5 plays an important role in the osteogenic differentiation of hADSCs through the modulation of the NF-${\kappa}B$ pathway.

Mesenchymal stem cells and osteogenesis

  • Jung, Cho-Rok;Kiran, Kondabagil R.;Kwon, Byoung S.
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.179-186
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    • 2001
  • Bone marrow stroma is a complex tissue encompassing a number of cell types and supports hematopiesis, differentiation of erythreid, nyel and lymphoid lineages, and also maintains undifferentiated hematopoietic stem cells. Marrow-derived stem cells were composed of two populations, namely, hematopoietic stem cells that can differentiate into blood elements and mesenchymal stem cells that can give rise to connective tissues such as bone, cartilage, muscle, tendon, adipose and stroma. Differentiation requires environmental factors and unique intracellular signaling. For example, $TGF-{\beta}$ or BMP2 induces osteoblastic differentiation of mesenchymal stem are very exciting. However, the intrinsic controls involved in differentiation of stem cells are yet to be understood properly in order to exploit the same. This review presents an overview of the recent developments made in mesenchymal stem cell research with respect to osteogenesis.

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Methyltransferase and demethylase profiling studies during brown adipocyte differentiation

  • Son, Min Jeong;Kim, Won Kon;Oh, Kyoung-Jin;Park, Anna;Lee, Da Som;Han, Baek Soo;Lee, Sang Chul;Bae, Kwang-Hee
    • BMB Reports
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    • v.49 no.7
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    • pp.388-393
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    • 2016
  • Although brown adipose tissue is important with regard to energy balance, the molecular mechanism of brown adipocyte differentiation has not been extensively studied. Specifically, regulation factors at the level of protein modification are largely unknown. In this study, we examine the changes in the expression level of enzymes which are involved in protein lysine methylation during brown adipocyte differentiation. Several enzymes, in this case SUV420H2, PRDM9, MLL3 and JHDM1D, were found to be up-regulated. On the other hand, Set7/9 was significantly down-regulated. In the case of SUV420H2, the expression level increased sharply during brown adipocyte differentiation, whereas the expression of SUV420H2 was marginally enhanced during the white adipocyte differentiation. The knock-down of SUV420H2 caused the suppression of brown adipocyte differentiation, as compared to a scrambled control. These results suggest that SUV420H2, a methyltransferase, is involved in brown adipocyte differentiation, and that the methylation of protein lysine is important in brown adipocyte differentiation.

Extracts of Korean Medicinal Plant Extracts Alter Lipogenesis of Pig Adipose Tissue and Differentiation of Pig Preadipocytes In vitro (한국 약용식물 추출물이 In vitro 돼지 지방조직의 지방합성과 지방전구세포의 분화에 영향을 미친다)

  • Choi, Young-Suk;Choi, Kang-Duk;Kim, Sung-Do;Phillip, Owens;Chung, Chung-Soo
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.383-388
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    • 2010
  • Identification of natural compounds that can prevent the development of obesity in vivo is time consuming and expensive. We have used in vitro systems derived from pig adipose tissue to screen simple aqueous or ethanolic extracts of Korean medicinal herbs (KMH) for their anti-adipogenic potential. A total of 183 extracts were tested for their actions in lipogenesis of pig adipose tissue and differentiation of pig preadipocytes. Ethanol extracts were prepared from 72 and aqueous extracts were prepared from 111 medicinal herbs. Both an ethanolic and an aqueous extract were prepared from 65 of these. Thirteen extracts substantially altered rates of lipogenesis in vitro. The effects of KMH on lipogenesis of pig adipose tissue are as follows. Elevens reduced lipogenesis to rates that were more than 40% lower than control and four of these reduced rates of lipogenesis by more than 70%. The most potent anti-lipogenic extracts were those obtained in ethanol from Iridaceae and from Sophora flavescens AIT as well as both the aqueous and ethanolic extracts from Lysimachia vulgaris L. Two extracts, those prepared in water from Caesalpiniae lignum and from Phellodendri cortex, were found to promote rates of lipogenesis in vitro. The effects of KMH on differentiation of pig preadoipocytes are as follows. Twentyeight extracts altered the rates of differentiation of cultured porcine preadipocytes. Sixteen increased and twelve reduced the rates of differentiation of preadipocytes. Extracts prepared in ethanol from Moutan radicis cortex and from Ostericum koreanum and those prepared in water from Angelicae gigantis radix, from Inula henenium L and from Magnolia flos doubled the rate of differentiation of cultured porcine preadipocytes. Ten extracts reduced the in vitro rate of differentiation of porcine preadipocytes by more than 35%. These were the ethanolic extracts from Glycyrrizae radix, Nepetae spica and from Polygala myrtifolia and the aqueous extracts from Amaranthaceae, Asparagus cochinchinesis, Atractylodis rhizoma alba, Citrus junos TANAKA, Cyperus rotundus, Epimedium grandiflorum and from Moutan radicis cortex. Only the ethanolic extract from Polygala myrtifolia was able to both reduce lipogenesis in adipose tissue slices and retard differentiation of cultured preadipocytes. The results of our study will provide meaningful information to identify medicinal herbs which would reduce fat deposition in livestocks and humans.