• Title/Summary/Keyword: Cultured bone cell

Search Result 292, Processing Time 0.021 seconds

Regulation of Immune Responses by the Activating and Inhibitory Myeloid-Associate Immunoglobuline-Like Receptors (MAIR) (CD300)

  • Shibuya, Akira;Nakahashi-Oda, Chigusa;Tahara-Hanaoka, Satoko
    • IMMUNE NETWORK
    • /
    • v.9 no.2
    • /
    • pp.41-45
    • /
    • 2009
  • Activating and inhibitory cell surface receptors play important roles in regulation of immune responses. Recent progress has demonstrated that many inhibitory receptors pair with activating, as well as inhibitory, isoforms, both of whose genes are located in small clusters on a chromosome. We and others identified paired activating and inhibitory immunoglobulin-like receptors, designated myeloid-associated immunoglobulin-like receptors (MAIR) (CD300). MAIR is a multigene family consisting of nine genes on a small segment of mouse chromosome 11. MAIR family receptors are preferentially expressed on myeloid cells, including macrophages, dendritic cells, granulocytes, and bone-marrow-derived cultured mast cells, and a subset of B cells and regulate activation of these cells. Thus, MAIR plays an important role in innate immunity mediated by myeloid cells.

Effect of Vitamin D3 on Biosynthesis of Estrogen in Porcine Granulosa Cells via Modulation of Steroidogenic Enzymes

  • Hong, So-Hye;Lee, Jae-Eon;An, Sung-Min;Shin, Ye Young;Hwang, Dae Youn;Yang, Seung Yun;Cho, Seong-Keun;An, Beum-Soo
    • Toxicological Research
    • /
    • v.33 no.1
    • /
    • pp.49-54
    • /
    • 2017
  • Vitamin D3 is a fat-soluble secosteroid responsible for enhancing intestinal absorption of calcium, iron, and other materials. Vitamin D3 deficiency, therefore, can cause health problems such as metabolic diseases, and bone disorder. Female sex hormones including estrogen and progesterone are biosynthesized mainly in the granulosa cells of ovary. In this study, we isolated granulosa cells from porcine ovary and cultured for the experiments. In order to examine the effect of vitamin D3 on the ovarian granulosa cells, the mRNA and protein levels of genes were analyzed by real-time PCR and Western blot assay. The production of estrogen from the granulosa cells was also measured by the ELISA assay. Genes associated with follicle growth were not significantly altered by vitamin D3. However, it increases expression of genes involved in the estrogen-biosynthesis. Further, estrogen concentrations in porcine granulosa cell-cultured media increased in response to vitamin D3. These results showed that vitamin D3 is a powerful regulator of sex steroid hormone production in porcine granulosa cells, suggesting that vitamin D deficiency may result in inappropriate sexual development of industrial animals and eventually economic loss.

HSP27 EXPRESSION IN OSTEOBLAST BY THERMAL STRESS (골모세포에서 열자극에 의한 Hsp27 발현에 대한 연구)

  • Rim, Jae-Suk;Kim, Byeong-Ryol;Kwon, Jong-Jin;Jang, Hyon-Seok;Lee, Eui-Suk;Jun, Sang-Ho;Woo, Hyeon-Il
    • Maxillofacial Plastic and Reconstructive Surgery
    • /
    • v.30 no.1
    • /
    • pp.11-21
    • /
    • 2008
  • Aim of the study: Thermal stress is a central determinant of osseous surgical outcomes. Interestingly, the temperatures measured during endosseous surgeries coincide with the temperatures that elicit the heat shock response of mammalian cells. The heat shock response is a coordinated biochemical response that helps to protect cells from stresses of various forms. Several protective proteins, termed heat shock proteins (hsp) are produced as part of this response. To begin to understand the role of the stress response of osteoblasts during surgical manipulation of bone, the heat shock protein response was evaluated in osteoblastic cells. Materials & methods: With primary cell culture studies and ROS 17/2.8 osteoblastic cells transfected with hsp27 encoding vectors culture studies, the thermal stress response of mammalian osteoblastic cells was evaluated by immunohistochemistry and western blot analysis. Results: Immunocytochemistry indicated that hsp27 was present in unstressed osteoblastic cells, but not fibroblastic cells. Primarily cultured osteoblasts and fibroblasts expressed the major hsp in response to thermal stress, however, the small Mr hsp, hsp27 was shown to be a constitutive product only in osteoblasts. Creation of stable transformed osteoblastic cells expressing abundant hsp27 protein was used to demonstrate that hsp27 confers stress resistance to osteoblastic cells. Conclusions: The demonstrable presence and function of hsp27 in cultured bones and cells implicates this protein as a determinant of osteoblastic cell fate in vivo.

Nanotechnology in the Surface Treatment of Titanium Implant. (임상가를 위한 특집 2 - 티타늄 임플란트 표면처리에서의 나노테크놀로지)

  • Oh, Seung-Han
    • The Journal of the Korean dental association
    • /
    • v.48 no.2
    • /
    • pp.106-112
    • /
    • 2010
  • Tissue engineering has been enhanced by advance in biomaterial nature, surface structure and design. In this paper, I report specifically vertically aligned titania ($TiO_2$) nanotube surface structuring for optimization of titanium implants utilizing nanotechnology. The formation, mechanism, characteristics of titania nanotubes are explained and emerging critical role in tissue engineering and regenerative medicine is reviewed. The main focus of this paper is on the unique 3 dimensional tubular shaped nanostructure of titania and its effects on creating epochal impacts on cell behavior. Particularly, I discuss how different cells cultured on titania nanotube are adhered, proliferated, differentiated and showed phenotypic functionality compared to those cultured on flat titanium. As a matter of fact, the presence of titania nanotube surface structuring on titanium for dental applications had an important effect improving the proliferation and mineralization of osteoblasts in vitro, and enhancing the bone bonding strength with rabbit tibia over conventional titanium implants in vivo. The nano-features of titania nanotubular structure are expected to be advantageous in regulating many positive cell and tissue responses for various tissue engineering and regenerative medicine applications.

Biological Effects of Ceramic-coating on Titanium

  • Sohn, Sung-Hwa;Lee, Jae-Bum;Kim, Ki-Nam;Kim, Hye-Won;Kim, In-Kyoung;Lee, Seung-Ho;Seo, Sang-Hui;Kim, Yu-Ri;Lee, Seung-Min;Shin, Sang-Wan;Ryu, Jae-Jun;Kim, Meyoung-Kon
    • Molecular & Cellular Toxicology
    • /
    • v.2 no.2
    • /
    • pp.97-105
    • /
    • 2006
  • Several features of the implant surface, such as roughness, topography, and composition play a relevant role in implant integration with bone. This study was conducted in order to determine the effects of ceramic-coatings on Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on Zr (Zrconium-coated surface), Nb (Niobium-coated surface), and control (Uncoated Titanium) Ti. The morphology of these cells was assessed by SEM. The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1,152 elements). The appearances of the surfaces observed by SEM were different on each of the three dental substrate types. MG63 cells cultured on Zr, Nb and control exhibited cell-matrix interactions. In the expression of several genes were up-, and down-regulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface morphology of the dental materials used.

Gene Expression of Osteosarcoma Cells on Various Coated Titanium Materials

  • Sohn, Sung-Hwa;Lee, Jae-Bun;Kim, Ki-Nam;Kim, In-Kyoung;Lee, Seung-Ho;Kim, Hye-Won;Seo, Sang-Hui;Kim, Yu-Ri;Shin, Sang-Wan;Ryu, Jae-Jun;Kim, Meyoung-Kon
    • Molecular & Cellular Toxicology
    • /
    • v.3 no.1
    • /
    • pp.36-45
    • /
    • 2007
  • Several features of the implant surface, such as topography, roughness, and composition play a relevant role in implant integration with bone. This study was conducted in order to determine the effects of different-coatings on Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on HA (Hydroxyapatite coating on Titanium), Ano (HA coating on anodized surface Titanium), Zr (zirconium-coating on Titanium), and control (non-coating on Titanium). The morphology of these cells was assessed by SEM. The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1,152 elements). The appearances of the surfaces observed by SEM were different on each of the three dental substrate types. MG63 cells cultured on HA, Ano, Zr, and control exhibited cell-matrix interactions. In the expression of several genes were up-, and down-regulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface morphology of the dental materials used.

Isolation of Mesenchymal Stem-like Cells from a Pituitary Adenoma Specimen

  • Shim, Jin-Kyoung;Kang, Seok-Gu;Lee, Ji-Hyun;Chang, Jong Hee;Hong, Yong-Kil
    • Biomedical Science Letters
    • /
    • v.19 no.4
    • /
    • pp.295-302
    • /
    • 2013
  • Some of the pituitary adenomas are invasive and spread into neighboring tissues. In previous studies, the invasion of pituitary adenomas is thought to be associated with epithelial-mesenchymal transition (EMT). In addition to that, we thought that mesenchymal stem cells (MSCs) exist in relevant microenvironment in pituitary adenoma. However, it has been little known about the existence of MSCs from pituitary adenoma. So we investigated whether mesenchymal stem-like cells (MSLCs) can be isolated from the pituitary adenoma specimen. We isolated and cultured candidate MSLCs from the fresh pituitary adenoma specimen with the same protocols used in culturing bone marrow derived MSCs (BM-MSCs). The cultured candidate MSLCs were analyzed by fluorescence-activated cell sorting (FACS) for surface markers associated with MSCs. Candidate MSLCs were exposed to mesenchymal differentiation conditions to determine the mesenchymal differentiation potential of these cells. To evaluate the tumorigenesis of candidate MSLCs from pituitary adenoma, we implanted these cells into the brain of athymic nude mice. We isolated cells resembling BM-MSCs named pituitary adenoma stroma mesenchymal stem-like cells (PAS-MSLCs). PAS-MSLCs were spindle shaped and had adherent characteristics. FACS analysis identified that the PAS-MSLCs had a bit similar surface markers to BM-MSCs. Isolated cells expressed surface antigen, positive for CD105, CD75, and negative for CD45, NG2, and CD90. We found that these cells were capable of differentiation into adipocytes, osteocytes and chondrocytes. Tumor was not developed in the nude mice brains that were implanted with the PAS-MSLCs. In this study, we showed that MSLCs can be isolated from a pituitary adenoma specimen which is not tumorigenic.

A Study on Anti-Bone Resorption & Osteoporosis by Taeyoungion-Jahage Extracts

  • Bae, Hyo-Sang;Cho, Hyung-Lae;Kim, Dong-Il;Lee, Tae-Kyun;Kim, Jun-Ki;Shin, Jung-Sik
    • The Journal of Korean Obstetrics and Gynecology
    • /
    • v.15 no.4
    • /
    • pp.61-75
    • /
    • 2002
  • Recombinant human $interleukin-1{\beta}$ $(rhIL-1{\beta})$ regulates several activities of the osteoblast cells derived from mouse calvarial bone explants in vitro. $rhIL-1{\beta}$ stimulated cellular proliferation and the synthesis of prostaglandin $E_2(PGE_2)$ and plasminogen activator activity in the cultured cells in a dose-dependent manner. However, the induction of osteocalcin synthesis and alkaine phosphatase activity in response to vitamine D, two characteristics of the osteoblast phenotype, were antagonized by $rhIL-1{\beta}$ over a similar dose range. This study supports the role of $IL-1{\beta}$ in the pathological modulation of bone cell metabolism, with regard to implication in the pathogenesis of osteoporosis by $IL-1{\beta}$. When the mouse calvarial bone cells were used, the bone resorption induced by $IL-1{\beta}$ was strongly inhibited by calcitonin treatment, indicating osteoclast-mediated bone resorption. On the other hand, the medicinal extracts of Taeyoungjon-Jahage (T.Y.J-J.H.G extracts) was tested for whether they could inhibit $IL-1{\beta}-induced$ $PGE_2$ production. Cell viability was not significantly affected by treatment with the indicated concentration of the extracts. The T.Y.J.-J.H.G. extracts were shown to have the inhibitory effects against the synthesis of $PGE_2$. We also examined the effect of the pretreatment with a various concentrations of the T.Y.J.-J.H.G. extracts then treated the $PGE_2-induction$ agents. Pretreatment of the T.Y.J.-J.H.G. extracts for 1 h, which by itself had little effect on cell survival, did not enhance the synthesis of $PGE_2$. Furthermore, the T.Y.J-J.H.G. extracts were shown to have the protective effects against plasminogen dependent fibrinolysis induced by the bone resorption agents of $IL-1{\beta}$. Pretreatment of the T.Y.J.-J.H.G. extracts for 1 h did not enhance the plasminogen dependent fibrinolysis. Finally, calcitonin showed the inhibitory activity the $IL-1{\beta}-stimulated$ bone resorption in the mouse calvarial bone cells having both of the osteoblast and osteoclast cells. Seemingly, pretreatment of the T.Y.J.-J.H.G. extracts for 1 h reduced the bone resorption. These results clearly indicated that calcitonin and T.Y.J.-J.H.G. extracts play key roles in inhibition of the osteoclast-mediated bone resorption.

  • PDF

Human Cord Serum as a Fetal Bovine Serum Substitute for the Culture of Human Amnion-Derived Stem Cells (인간의 양막유래 줄기세포의 체외 배양 시 소태아혈청 대체제로서의 인간제대혈청)

  • Kim, Jin-Young;Park, Se-A;Kang, Hyun-Mi;Kim, Eun-Su;Kim, Hae-Kwon
    • Development and Reproduction
    • /
    • v.11 no.2
    • /
    • pp.85-96
    • /
    • 2007
  • Mesenchymal stem cells (MSC) are promising candidates for cell-based therapies. One major obstacle for their clinical use is the unsafety of fetal bovine serum (FBS), which is a crucial part of all media currently used for the culture of MSC. We investigated the effect of human cord serum (HCS) on the growth response, mRNA and protein expressions of human amnion-derived stem cells (HAM). HAM were isolated from the amnion after a Caesarean section and cultured in DMEM supplemented with 10% FBS, 5% HCS or 10% HCS. During culture, their biological characteristics at earlier and later passages were analyzed using RT-PCR and immunocytochemistry. Regardless of serum sources, HAM showed the prominent expression of Oct-4, Rex-1, SCF, FGF-5, BMP-4, nestin, GATA-4, NCAM and HLA ABC genes. The expression profile was observed even at later passages. Similarly, HAM cultured in either FBS or HCS exhibited the distinct protein expression of collagen I, II, III and XII, fibronectin, $\alpha$-smooth muscle actin, vimentin, CK18, CD54, FSP, TRA-1-60, SSEA-3, -4 and HLA ABC. However, desmin expression was only observed in HAM cultured in medium supplemented with FBS and vWF expression was only found in HAM cultured in medium supplemented with HCS. Overall pattern of gene and protein expression of HAM was typical of known adult stem cells such as bone marrow-derived MSC. In conclusion, HCS could be as effective as FBS for the culture of HAM.

  • PDF

Expression of Human Growth Hormone Gene using Retrovirus Vector System In Vitro (In Vitro에서 Retrovirus Vector System을 이용한 인간 성장 호르몬 유전자의 발현)

  • Kim, Min-Ki;Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
    • /
    • v.35 no.3
    • /
    • pp.251-256
    • /
    • 2011
  • Human growth hormone (hGH), one of the most important hormones in medicine, is secreted from anterior pituitary gland. Its broad physiological function includes body growth, cell regeneration, increasement of muscle volume, bone density, body fat reduction, and so on. Due to the wide range of therapeutic effects, the hGH produced from E. coli has been commercialized already. In this study, we asked whether it is possible to produce recombinant hGH efficiently from various cultured mammalia cells. To meet this purpose, we chose a retrovirus vector system for transfer and expression of the hGH gene in various mammalian cells. Analyses of RT-PCR, ELISA, and Western blot to determine expression of the hGH gene showed the highest production of the hGH was determined from chicken embronic fibroblast (CEF) cells with the concentration of 8.58 ${\mu}g$/ml. The biological activity of the hGH was similar to the commercially available counterpart. These results suggest that mass production of hGH is possible not only in the E. coli but also in the various mammalian cells.