• Title/Summary/Keyword: in vitro Differentiation

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ISOLATION OF HUMAN ALVEOLAR BONE-DERIVED CELLS AND IN VITRO AMPLIFICATION FOR TISSUE ENGINEERING (조직공학용 사람 치조골세포의 인공증식)

  • Choi, Byung-Ho;Park, Jin-Hyoung;Huh, Jin-Young;Yoo, Jae-Ha
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.5
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    • pp.453-456
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    • 2001
  • Background: Autogenous alveolar bone cell transplantation may be suitable for tissue engineering for alveolar bone reconstruction. This study aimed to isolate human alveolar bone-derived cells (HABDCs) and to evaluate the ability of collagen gels to support HABDC proliferation and differentiation for human alveolar bone tissue engineering applications. Method: Cultures of primary HABDCs were established from alveolar bone chips obtained from 10 persons undergoing tooth extraction. These cells were expanded in vitro until passage 3 and used for the in vitro characterization of HABDCs and the in vitro analysis of collagen gels for alveolar bone tissue engineering. Results: Of the 10 attempts made to obtain HABDC cultures, eight were successful. HABDCs expressed the osteoblastic phenotype characterized by alkaline phosphatase activity, osteocalcin expression and the mineralization of the extracellular matrix in vitro. When seeded on collagen gels, HABDCs penetrated into the collagen gel matrices and proliferated inside the gels. Significantly, when HABDCs were embedded into the gels, collagen fibers and mineralization were produced within the gels. Conclusion: This study demonstrates the feasibility of using cultured HABDCs and collagen gels for human alveolar bone tissue engineering applications.

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The Anti-obesity Effect of Seungyangjeseup-tang for High Fat Diet Induced Obese Mice (고지방식이 유도 비만 생쥐에 대한 승양제습탕의 항비만 효과)

  • Kim, Jung-Min;Choi, Soo-Min;Woo, Chang-Hoon;Ahn, Hee-Duk
    • Journal of Korean Medicine Rehabilitation
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    • v.28 no.3
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    • pp.1-11
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    • 2018
  • Objectives This study was designed to evaluate the efficacy of Seungyangjeseup-tang on obesity by using 3T3-L1 cells and high fat diet mice. Methods In vitro, Seungyangjeseup-tang extract (SYJST) (10, 50, 100, 200, 400, $800{\mu}g/mL$) ware added in 3T3-L1 cells. SYJST cytotoxicity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assasy. Adipocyte differentiation was measured by Oil Red-O staining. In vivo, the experimental animals were divided into five groups: normal diet-fed normal group (N), high-fat diet (HFD)-fed control group (Con), HFD+SYJST 100 mg/kg group (SY100), HFD+SYJST 200 mg/kg group (SY200), and HFD+olistat 60 mg/kg as a positive drug control group (Orli). Markers of obesity, such as body weight, liver weight, food intake, serum total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C), liver tissue TC, TG and fecal TC, TG were measured. Results In vitro, cytotoxicity was not significant compared with the control group. 3T3-L1 cell's differentiation was significantly decreased in Oil Red-O staining. In vivo, compared with controls, mice treated with SYJST demonstrate lower body and liver weight, and reduced food intake. In addition, SYJST increased TC, TG in the serum but not significance. And SYJST showed decreasing tendency TC, TG in the liver tissue. Furthermore, SYJST increased TC, TG in the facal but not significance. Conclusions Based on the results above, Seungyangjeseup-tang may reduce adipocyte differentiation, body fat, food intake, liver weight in obesity. This suggests that Seungyangjeseup-tang may be clinically useful in obesity treatment.

Toxoplasmacidal Effect of HL-60 Cells Differentiated by Dimethylsulfoxide (Dimethylsulfoxide로 분화시킨 HL-60 세포의 yoxoplasma 파괴 효과)

  • Choe, Won-Yeong;Nam, Ho-U;Yu, Jae-Eul
    • Parasites, Hosts and Diseases
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    • v.26 no.4
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    • pp.229-238
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    • 1988
  • In vitro culture of Toxoplasma gondii in HL-60 cells and cell-mediates immunity against Toxoplasma in dimethylsulfoxide(DMSO) -induced HL-60 cells, i.e., differentiation into granulocytes, were pursued. HL-60 calls were treated with various concentrations of DMSO, and 1.3%(v/v) for 3 day incubation was chosen as the optimal condition icy differentiation into granulocytes. The degree of differentiation was assayed in physiological and functional aspects in addition to morphological point. When treated with 1.3% DMSO for 3 days, HL-60 cells did not synthesiar DNA materials beyond background level, and showed active chemotactic response to chemotactic peptide, formal-methionyl-leucyl-phenylalanine(FMLP). Morphologically promyelocytes of high nuclearlcytoplasmic(NIC) ratio changed to granulocytes of relatively low WJC ratio. The relationships between HL-60 cells or DMSO-induced HL-60 cells and Toxoplasma were examined after stain with Giemsa and Buorescent dye (acridine orange). HL-60 cells did not show any sign of torso- plasmacidal activity but showed intracellular proliferation of Texoplasma to form rosette for 72 hr co-culture. In contrast, OMSO-induced HL-60 cells phagocytosed Toxoplasma within 1 hr, and performed a process of intracellular digestion of Toxoplasma thereafter. With the above results, it is suggested that phagosome-Iysosome fusion is one of the critical events for the parasitism by Toxoplasma or for susceptibility of host cells. The in vitro culture system of this study has offered a defined condition to study the protozoan parasite-host cell interactions.

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Differentiation and Labeling of Mouse Preadipocytes for Allogenic Transplantation Study (동종이식 연구를 위한 마우스 지방전구세포의 표지 및 분화 방법의 확립)

  • Kim, In Ok;Kim, Taek Seung;Kim, Mi Hyung;Hyon, Won Sok;Mun, Goo Hyun;Oh, Kap Sung;Bang, Sa Ik
    • Archives of Plastic Surgery
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    • v.32 no.4
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    • pp.533-538
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    • 2005
  • Due to its safety and softness, autologous fat transplantation has been commonly performed for soft tissue correction. However, the injected fat is absorbed resulting in the reduction of volume of the graft by 40-60% within a few months. Thus, there was an attempt to use adipocytes differentiated from preadipocytes in vitro for transplantation. Differentiated adipocytes were biocompatible and matured with gradual volume increase at transplantation site in clinical study(unpublished data). In addition, they did not induce immune rejection in response to nonself lymphocytes in a mixed lymphocyte reaction(MLR)(unpublished data). The purpose of this study is to differentiate mouse preadipocytes following labeling into adipocytes to establish an animal model for allogenic transplantation. Preadipocytes isolated from inguinal and retroperitoneal fat pad of C57BL/6 mice were proliferated with growth medium by passage 3 and differentiated into adipocytes with different culture conditions after labeled with BrdU. At most suitable conditions, above 90% of preadipocytes were differentiated and BrdU labeling did not affect differentiation rate and function of differentiated adipocytes. These results demonstrate that BrdU-labeled adipocytes resulting from this in vitro differentiation protocol are useful for allogenic transplantation study.

Suppression of HIF-1α by Valproic Acid Sustains Self-Renewal of Mouse Embryonic Stem Cells under Hypoxia In Vitro

  • Lee, Hyo-Jong;Kim, Kyu-Won
    • Biomolecules & Therapeutics
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    • v.20 no.3
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    • pp.280-285
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    • 2012
  • The developing embryo naturally experiences relatively low oxygen conditions in vivo. Under in vitro hypoxia, mouse embryonic stem cells (mESCs) lose their self-renewal activity and display an early differentiated morphology mediated by the hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$). Previously, we demonstrated that histone deacetylase (HDAC) is activated by hypoxia and increases the protein stability and transcriptional activity of HIF-$1{\alpha}$ in many human cancer cells. Furthermore HDAC1 and 3 mediate the differentiation of mECSs and hematopoietic stem cells. However, the role of HDACs and their inhibitors in hypoxia-induced early differentiation of mESCs remains largely unknown. Here, we examined the effects of several histone deacetylase inhibitors (HDACIs) on the self-renewal properties of mESCs under hypoxia. Inhibition of HDAC under hypoxia effectively decreased the HIF-$1{\alpha}$ protein levels and substantially improved the expression of the LIF-specific receptor (LIFR) and phosphorylated-STAT3 in mESCs. In particular, valproic acid (VPA), a pan HDACI, showed dramatic changes in HIF-$1{\alpha}$ protein levels and LIFR protein expression levels compared to other HDACIs, including sodium butyrate (SB), trichostatin A (TSA), and apicidin (AP). Importantly, our RT-PCR data and alkaline phosphatase assays indicate that VPA helps to maintain the self-renewal activity of mESCs under hypoxia. Taken together, these results suggest that VPA may block the early differentiation of mESCs under hypoxia via the destabilization of HIF-$1{\alpha}$.

Induction of Differentiation on the Human Histocytic Lymphoma Cell Line U-937 by Costunolide (Costunolide의 백혈병 세포주 U-937에 대한 분화 유도 작용)

  • Kim, Joo-Il;Lee, Sung-Ho;Park, Jae-Hoon;Park, Hee-Joun;Lee, Kyung-Tae
    • Korean Journal of Pharmacognosy
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    • v.30 no.1
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    • pp.7-11
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    • 1999
  • The present work was carried out to examine the effect of costunolide on the growth of several cells and characteristics of U-937 human leukemia-derived cell line. Costunolide produced a potent antitumor activity in vitro dependent on concentration against several tumor cells such as P-388, L-1210 leukemia and SNU-5 stomach cancer cells. However, it showed less cytotoxicity on normal cells such as Maccaccus rheus monkey kidney cells (MA-104) up to 200 ${\mu}M$ concentration. An effect of cell differentiation by costunolide was assessed by its ability to reduce nitroblue tetrazolium (NBT), and to induce phagocytosis of latex particles. In order to establish whether costunolide induces U-937 cells to differentiate toward macrophage or granulocyte, esterase activities was measured. Based on these results, we found that costunolide having cytotoxicity on U-937 human leukemia cells was explained through differentiation inducing activity.

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Osteoblast adhesion and differentiation on magnesium titanate surface (마그네슘 티타네이트 표면의 조골세포 부착도와 분화)

  • Choi, Seung-Min;Lee, Jae-Kwan;Ko, Sung-Hee;Um, Heung-Sik;Chang, Beom-Seok
    • Journal of Periodontal and Implant Science
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    • v.35 no.4
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    • pp.851-861
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    • 2005
  • The nature of the implant surface can directly influence cellular response, ultimately affecting the rate and quality of new bone tissue formation. The aim of this in vitro study was to investigate if human osteoblast-like cells, Saos-2, would respond differently when plated on disks of magnesium titanate and machined titanium. Magnesium titanate disks were prepared using Micro Arc Oxidation(MAO) methods. Control samples were machined commercially pure titanium disks. The cell adhesion, proliferation and differentiation were evaluated by measuring cell number, and alkaline phosphatase(ALPase) activity at 1 day and 6 day after plating on the titanium disks. Measurement of cell number and ALPase activity in Saos-2 cells at 1 day did not demonstrate any difference between machined titanium and magnesium titanate. When compared to machined titanium disks, the number of cells was reduced on the magnesium titanate disks at 6 day, while ALPase activity was more pronounced on the magnesium titanate. Enhanced differentiation of cells grown on magnesium titanate samples was indicated by decreased cell proliferation and increased ALPase activity.