• Title/Summary/Keyword: FGF-2

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Maintenance of Proliferation and Adipogenic Differentiation by Fibroblast Growth Factor-2 and Dexamethasone Through Expression of Hepatocyte Growth Factor in Bone Marrow-derived Mesenchymal Stem Cells

  • Oh, Ji-Eun;Eom, Young Woo
    • Biomedical Science Letters
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    • v.22 no.1
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    • pp.1-8
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    • 2016
  • Several studies have investigated the various effects of dexamethasone (Dex) on the proliferation and differentiation of mesenchymal stem cells (MSCs). Previously, we reported that co-treatment with L-ascorbic acid 2-phosphate and fibroblast growth factor (FGF)-2 maintained differentiation potential in MSCs through expression of hepatocyte growth factor (HGF). In this study, we investigated the effects of co-treatment with FGF-2 and Dex on the proliferation and differentiation potential of MSCs during a 2-month culture period. Co-treatment with FGF-2 and Dex increased approximately a 4.7-fold higher accumulation rate of MSC numbers than that by FGF-2 single treatment during a 2-month culture period. Interestingly, co-treatment with FGF-2 and Dex increased expression of HGF and maintained adipogenic differentiation potential during this culture period. These results suggest that co-treatment with FGF-2 and Dex preserves the proliferation and differentiation potential during long-term culture.

Role of FGF and MEK Signaling in Formation of the Hydrostatic Pressure Receptor Cells during Ascidian Embryogenesis (멍게의 수압수용체세포 형성에서 FGF와 MEK 신호의 역할)

  • Seo, Hyeong-Joo;Kim, Gil-Jung
    • Development and Reproduction
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    • v.13 no.4
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    • pp.291-296
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    • 2009
  • In most larvae of ascidian, two sensory pigment cells, otolith and ocellus, lie in their brain vesicle. They also have a third type of sensory cells: hydrostatic pressure receptor (Hpr) cells. The Hpr cells were presumed to be hydrostatic pressure-detection cells, but their precise functions is still disputed. In this study, we investigated whether an FGF signaling is involved in formation of Hpr cells. When fertilized eggs were injected with Hr-FGF9/16/20 antisense MO, the resulting larvae showed severe abnormalities with no expression of the Hpr cell-specific Hpr-1 antigen. Similar results were obtained using an FGF receptor inhibitor, SU5402, and an MEK inhibitor, U0126. Embryos treated with SU5402 or U0126 during the 32-cell and hatching stages did not express the Hpr-1 antigen. To elucidate the temporal requirement for the FGF signaling in formation of Hpr cells, embryos were treated with SU5402 for 2 h, or U0126 for 20 min during various embryonic stages. Larvae treated with SU5402 from the 16-cell stage to the 64-cell stage did not express the Hpr-1 antigen, whereas those treated at the early gastrula stage expressed the Hpr-1 antigen. When U0126 treatment was carried out at various stages between the 8-cell and late gastrula stages, larvae scarcely formed the Hpr cells. They showed expression of the Hpr-1 antigen when embryos were placed in U0126 just before the neural plate stage. These results suggest that FGF9/16/20 signaling is involved in formation of Hpr cells from the primary neural induction stage to the late gastrula stage.

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Effect of Fucoidan on Angiogenesis and Gene Expression in Human Umbilical Vein Endothelial Cells (후코이단이 혈관 내피세포의 신생혈관 생성 효과 및 관련 유전자의 발현에 미치는 영향)

  • Park, Ho;Kim, Beom-Su
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.323-328
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    • 2017
  • Angiogenesis is a process including members of the angiogenic factors. In particular, fibroblast growth factor 2 (FGF2) is considered the most potent angiogenic factor because it promotes cell proliferation and tube formation. A recent study reported that fucoidan derived from marine plant potentiated FGF-2 induced tube formation in human endothelial cells. On the other hand, the molecular mechanisms involved in the angiogenic activity of fucoidan and FGF2 are unknown. In this study, a fucoidan treatment promoted angiogenesis induced by FGF2. The effects of fucoidan on FGF2-induced angiogenesis were confirmed by a proliferation assay using a CellTiter96 Aqueous One solution after a treatment with fucoidan and FGF2. The tube formation and wound healing assay for the angiogenic activity were also confirmed. Reverse transcription PCR showed a change in the mRNA of vascular endothelial growth factor-A (VEGF-A), intercellular adhesion molecule-1 (ICAM-1), matrix metallopeptidase9 (MMP9), and the signal transducer and activator of transcription3 (STAT3). In summary, the Fucoidan/FGF2 treatment induced an increase in cell proliferation, improved the tube formation and wound healing activity, and altered the STAT3, VEGF-A, ICAM-1, and MMP9 mRNA expression levels. Further research will be needed to provide a scientific explanation in terms of cell-signaling and confirm the present findings.

Possible Implication for an Indirect Interaction between Basic Fibroblast Growth Factor and (Na,K)ATPase

  • Oh, Ji-Hyun;Lee, Kyung-Lim
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.707-711
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    • 1998
  • The (Na,K)ATPase is responsible for generating the ionic gradients and membrane potentials by the exchange of intracellular $Na^+$ for $K^+$. It has been recentl y shown that (Na,K)ATPase is involved in the exocytic pathway of basic fibroblast growth factor (bFGF), although it is not known that bFGF is secreted to the outside of cell through direct interaction with (Na,K) ATPase. To understand the role for (Na,K)ATPase in the secretary pathway of bFGF, we have sought to identify the cytoplasmic domains of the alpha1 isoform of (Na,K)ATPase interacting with bFGF by yeast two-hybrid system. We have also investigated the interaction between the alpha2 isoform of (Na,K)ATPase and bFGF to find out whether the interaction is isoform-specific. We found that none of the cytoplasmic domains of (Na,K)ATPase isoforms interacted with bFGF. The result suggests that the interaction between bFGF and (Na,K)ATPase might be indirect, thus requiring other proteins which are involved in the formation of protein complexes for the interaction, although we cannot exclude the possibility that the interaction requires the element of the whole alpha subunit structure that was not present in the isolated alpha subunit cytoplasmic domains.

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The expression and function of FGF-8 in limb development and regeneration of mexican axolotl, Ambystoma mexicanum

  • Han, Man-Jong;Kim, Won-Sun
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.57-58
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    • 1998
  • From the present study, following conclusions can be drawn: 1. lide in other species, axolotl FGF-8 is proposed to play a similar role in the early phase of limb development. However, the mechanism of its expression might be somewhat different from amniotes considering its characteristic mesenchymal expression. 2. In the regenerating axolotl limbs, Fgf-8 expression profile suggests that it is involved in wound gealing, dedifferentiation, and blastema formation. 3. Exoggenously supplied FGF-8 can accelerate blastema formation and concomitantly increase the Msx-1 expression level at the early stage of limb regeneration. Furthermore, it can partially substitute for nerve factor(s) as has been indicated by the induction of blastema formation in the denervated regenerates after FGF-8 application. 4. The unique expression feature of Fgf-8 in hte mesenechymal tissue of the regenerating axolotl limb might be casually related to its remarkable regeneration capacity of urodele.

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In Vitro Differentiation of Human Amniotic Membrane-derived Stem Cells into Hepatocyte-like Cells (양막 유래 줄기세포의 간세포로의 분화 유도)

  • Kook, Min-Ji;Park, Soo-Yeon;Kang, Hyun-Mi;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.10 no.1
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    • pp.63-73
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    • 2006
  • This study aimed to find out suitable culture conditions for the differentiation of human amniotic membrane-derived stem cells(HAM) into hepatocyte-like cells. Almost homogenous population of fibroblast-like cells was successfully isolated from the amniotic membrane. In comparison to the non-coated plates and in the absence of insulin/transferrin/selenium(ITS), HAM cultured on the fibronectin-coated plates and in the presence of ITS showed the more intense immunocytochemical staining against the albumin. Addition of both fibroblast growth factor(FGF)-1 and -2 to the differentiation medium gave stronger staining compared to the treatment with FGF-1 or -2 alone. Periodic acid Schiff's base staining of glycogen and morphological turnover of fibroblast-like appearance of HAM into round shape matched the results of immunocytochemical studies. When the efficiency of two-step culture method was examined on the differentiation of HAM into hepatocyte-like cells, all of the results of immunocytochemical staining, periodic acid Schiff's staining and morphological change exhibited effective hepatic differentiation of HAM compared to the continuous culture method. Immunoblot analyses of HAM- conditioned media against the albumin showed that the culture of HAM in the presence of both ITS and fibronectin always gave a stronger staining intensity than those in the absence of them, and that the addition of ether mixture of FGF-4 and either FGF-2 or transforming growth $factor(TGF)-{\alpha}$ to the culture medium significantly enhanced the albumin secretion by HAM. Based on these observations, it is suggested that HAM could differentiate into hepatocyte-like cells under a culture condition consisting of fibronectin and ITS, and addition of FGF-4 with either one of FGF-2 or $TGF-{\alpha}$ could enhance the hepatic differentiation of HAM.

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IMMUNOHISTOCHEMICAL PROFILE OF BASIC FIBROBLAST GROWTH FACTOR(bFGF) IN GROWING RAT T-M JOINT (성장 중인 흰쥐 악관절의 basic fibroblast growth factor(bFGF) 분포에 관한 면역조직화학적 연구)

  • Kim, Jong-Ha;Lee, Sang-Chull
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.1
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    • pp.1-8
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    • 2001
  • This study was designed to localize the distribution of basic fibroblast growth factor(bFGF) in the developing rat condylar region and to elucidate the associated function of bFGF in the condyle development. The condyles of temporomandibular joint of Sprague-Dawley rats (27g of weight) were used. The tissues were examined with electron microscope and immunohistochemical method. The results were as follows: 1. The developing condylar region are divided in to 5 zones apparently: proliferative, maturation, hypertrophic, calcifying, and ossification zones. 2. The cells in the proliferative zone are condensed and have under-developed cell organells in the cytoplasm. This zone shows a strong immunoreactivity of bFGF. 3. The cells in the maturation zone are typical chondroblasts showing well-developed cell organells and round nucleus. The cartilaginous matrix does not show the immunoreactivity of bFGF, while the chondroblasts show the immunoreactivity. 4. The cells in the hypertrophic zone show hypertrophic change having the degenerated cell organelles and small nucleus. There are no immunoreactivity of bFGF in this zone except the nucleus and endoplasmic region showing mild immunoreactivity. 5, The cells in the calcifying zone show hypertrophic change and cell organelles are disappeared. The cells are surrounded by the calcified cartilaginous matrix. There are no immunoreactivity of bFGF in this zone except the endoplasmic region showing mild immunoreactivity. 6. In the zone of bone formation, chondroblasts are disappeared. Newly differentiated osteoblasts secreting osteoid around the calcified cartilaginous matrix. The bone marrow shows the immunoreactivity of bFGF, while the bone matrix does not show the immunoreactivity of bFGF.

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Effects of FGF on Embryonic Development In Vitro in Hanwoo COCs (한우 난구 복합체의 체외발생에 있어서 FGF(Fibroblast Growth Factor)가 미치는 영향)

  • Choi S.H.;Cho S.R.;Kim H.J.;Choe C.Y.;Han M.H.;Son D.S.;Chung Y.G.;H. Hoshi
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.157-162
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    • 2006
  • It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs (cumulus oocytes complexes) in vitro. Attempts had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of FGF (fibroblast growth factor) on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in HPM 199 (Inst. of Functional peptide, Japan) containing 0.1, 1 and 10 ng/ml FGF for 24 hr, maturation rates to metaphase II ($70.0{\sim}75.0%$) were significantly higher (p<0.05) than that of control group (0 ng/ml FGF, 37.5%). When matured COCs with FGF were cultured in maturation medium after in vitro fertilization, developmental rates to blastocysts were 9.5, 0 and 2.9%, respectively, compared to 25.0% of the control group (p<0.05). When the matured COCs with FGF were cultured in HPM 199 (IFP971, Inst. of Functional peptide, Japan) containing 10% FBS, 0.8% BSA or 0.1% PVA (polyvinyl alcohol), the blastocyst formation rates were 12.4, 12.8 and 8.5%, respectively, while the rates of matured COCs with FGF and cultured with IVMD and IVD (Inst. of Functional peptide, Japan) without serum were 38.4% and 34.8%, respectively (p<0.05). These results suggested that FGF is available for in vitro maturation of bovine COCs and is not suitable for in vitro development, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine embryo development.

EFFECTS OF bFGF AND PDGF-BB ON OSTEOBLAST DIFFERENTIATION OF BONE MARROW-DERIVED MESENCHYMAL STEM CELL IN RAT (bFGF, PDGF-BB가 백서 골수기원 간엽 줄기세포의 조직골세포 분화에 미치는 영향에 관한 연구)

  • Song, Gin-Ah;Choi, Jin-Young
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.6
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    • pp.495-505
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    • 2006
  • In this study we evaluate the effects of bFGF-BB and PDGF on in vitro proliferation, differentiation and mineralization of mesenchymal stem cells (MSCs) from rat. MSCs were prepared from the bone marrow of 6 or 7-week-old male rats with a technique previously described by Maniatopoulos et al. in 1988. Lineage differentiation to osteogenesis, chondrogenesis and adipogenesis were performed. At first, we characterized the cultured cell on passage 1, 3, 5, 7 with immunocytochemical staining using CD29, 44, 34, 45, ${\alpha}$-SMA and type I collagen. And to study the effects of bFGF and PDGF-BB on proliferation, differentiation and mineralization, we seeded the expanded cell at a density of 6 $6{\times}10^3\;cells/cm^2$ to 100-mm dish for evaluation of cell proliferation and MTT assay was carried out on day 2, 4, 7, 9. We also resuspended the cells with same density $(6{\times}10^3\;cells/cm^2)$ to 24 well plates for subculture. On the following day, the attached cells were exposed to 2.5ng/ml bFGF and/or 25ng/ml PDGF-BB daily during 5 days. The osteocalcin (OC) level was assessed and mineral contents were evaluated with alizarin red S staining on subculture day 2, 7, 14, 21. We identified the mesenchymal stem cell from the bone marrow derived cells of rat through their successful multi-differentiation and stable display of its phenotype. And bFGF and PDGF-BB showed the effect that inhibited osteoblastic differentiation and mineralization mildly in above concentration at in vitro culture. This study was supported by grant 04-2004-0120 from the Seoul National University Hospital Research Fund.

Growth Factors Upregulated by Uric Acid Affect Guanine Deaminase-Induced Melanogenesis

  • Nan-Hyung Kim;Ai-Young Lee
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.89-96
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    • 2023
  • Uric acid produced by guanine deaminase (GDA) is involved in photoaging and hyperpigmentation. Reactive oxygen species (ROS) generated by uric acid plays a role in photoaging. However, the mechanism by which uric acid stimulates melanogenesis in GDA-overexpressing keratinocytes is unclear. Keratinocyte-derived paracrine factors have been identified as important mechanisms of ultraviolet-induced melanogenesis. Therefore, the role of paracrine melanogenic growth factors in GDA-induced hypermelanosis mediated by uric acid was examined. The relationships between ROS and these growth factors were examined. Primary cultured normal keratinocytes overexpressed with wild type or mutant GDA and those treated with xanthine or uric acid in the presence or absence of allopurinol, H2O2, or N-acetylcysteine (NAC) were used in this study. Intracellular and extracellular bFGF and SCF levels were increased in keratinocytes by wild type, but not by loss-of-function mutants of GDA overexpression. Culture supernatants from GDA-overexpressing keratinocytes stimulated melanogenesis, which was restored by anti-bFGF and anti-SCF antibodies. Allopurinol treatment reduced the expression levels of bFGF and SCF in both GDA-overexpressing and normal keratinocytes exposed to exogenous xanthine; the exogenous uric acid increased their expression levels. H2O2-stimulated tyrosinase expression and melanogenesis were restored by NAC pretreatment. However, H2O2 or NAC did not upregulate or downregulate bFGF or SCF, respectively. Overall, uric acid could be involved in melanogenesis induced by GDA overexpression in keratinocytes via bFGF and SCF upregulation not via ROS generation.