• 제목/요약/키워드: in vitro Differentiation

검색결과 749건 처리시간 0.023초

미숙화기와 액아에 의한 네오레게리아의 기내 번식 (In Vitro Propagation of Neoregeria carorinae cv. Tricolor from Immature Flowers and Lateral Buds)

  • 정향영;박봉규;유창재
    • 식물조직배양학회지
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    • 제22권4호
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    • pp.223-227
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    • 1995
  • 네오게레리아의 대량번식을 위해 미숙화기와 액아를 배양하였으며, 배양에 적합한 재료 채취시기, 생장조절제 종류와 농도 및 배양방법이 부정아 형성 및 생장에 미치는 영향을 조사한 결과, 미숙화기는 화아분화 4주후에, 액아는 화아 분화 7주후에서 절취하여 배양하는 것이 부정아 형성률이 가장 높았다. 부정아 형성에 적합한 생장조절제 종류와 농도는 MS + BA 1 mg/L + BA 0.5 mg/L이었다. 고체배지가 액체배지보다 부정아 형성 및 생장에 더 효과적이었다. 발근에 적합한 생장조절제 종류와 농도는 NAA 1 mg/L이었다. 액체배지가 고체배지보다 신초의 발근 및 생장에 더 효과적이었다.

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A Mouse Thymic Stromal Cell Line Producing Macrophage-Colony Stimulating Factor and Interleukin-6

  • Lee, Chong-Kil;Kim, Jeong-Ki;Kim, Kyungjae;Han, Seong-Sun
    • Archives of Pharmacal Research
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    • 제23권3호
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    • pp.252-256
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    • 2000
  • A thymic stromal cell line, TFGD, was established from a thymic tumor mass developed spontaneously in p53 knock out mouse, and was found to produce cytokines that could induce bone marrow hematopoietic stem cells (HSCs) to differentiate into macrophages. The cytokines produced by the TFGD line were assessed by immunoassays. High level of macrophage-colony stimulating factor (M-CSF) and interleukin (IL)-6 was detected in the TFGD-culture supernatant, whereas granulocyte/macrophage-colony stimulating factor (GM-CSF), IL-3, IL-4, IL-5, IL-13, or interferon (IFN)-$\gamma$ was undetectable. Blocking experiments showed that anti-M-CSF monoclonal antibody could neutralize the differentiation-inducing activity shown by the TFGD-culture supernatant. Dot blot analysis of the total RNA isolated from the cultured fetal thymic stromal cells showed that M-CSF transcripts were expressed in the normal thymus. These observations, together with the earlier finding that M-CSF plus IL-6 is the optimal combination of cytokines for the induction of macrophage differentiation from HSCs in vitro, may indicate that thymic macrophages could be generated within the thymus by cytokines involving M-CSF.

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마황(麻黃)과 석고(石膏) 혼합물이 지방전구세포 분화와 흰쥐의 비만에 미치는 영향 (Effect of Mixed Extract of Ephedrae Sinica and Fibrosum Gypsum on Differentiation of Preadipocytes and Obesity of Rats)

  • 기영범;김대훈;양미성;강대희;김선종;최진봉
    • 한방재활의학과학회지
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    • 제24권3호
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    • pp.11-27
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    • 2014
  • Objectives This study was designed to evaluate the effects of Ephedra Sinica and Fibrosum Gypsum extract on obesity by using 3T3-L1 cells and high fat diet rats. Methods In vitro, Ephedra Sinica and Fibrosum Gypsum extract (50, 100, 200, $500{\mu}g/ml$) were added in 3T3-L1 cells. Cytotoxicity was measured by MTT assasy. Adipocyte differentiation was measured by Oil Red O staining, GPDH activity and $C/EBP{\alpha}$ protein expression. In vivo, Sprague-Dawley rats were divided into 5 groups : Normal diet group (Normal group), taken high fat diet and no treatment group (Control group), taken high fat diet and orally administered Ephedra Sinica and Fibrosum Gypsum extract daily (Group I: 50 mg/kg, Group II: 100 mg/kg, Group III : 200 mg/kg, oral). For 6 weeks of administration, body weight and the amount of food intake were measured once a week. After administration, blood analysis (AST, ALT, T-Bilirubin, BUN, RBC, Hb, HCT), serum lipid level (triglyceride, Total cholesterol, HDL, LDL), serum leptin level, epididymal adipose tissue weight and histological finding of liver were estimated. Results In vitro, The cytotoxicity was not significant. 3T3-L1 cell's differentiation was significantly decreased in Oil Red O staining, GPDH activity and $C/EBP{\alpha}$ protein expression. In vivo, Body weight and the amount of food intake, AST, ALT, Total cholesterol, TG, LDL, serum leptin, epididymal adipose tissue weight showed significant decrease in group I, group II and group III. There were no significant difference in T-bilirubin, BUN, RBC, Hb and HCT between all groups. HDL showed significant increase in group I, group II and group III. In histological finding of liver tissue, there were decreased adiposity and cytopathic effect in group I, group II and group III. Conclusions It is suggested that Ephedra Sinica and Fibrosum Gypsum extract can be used in the treatment of obesity.

Role of CXCR7 and Effects on CXCL12 in SiHa Cells and Upregulation in Cervical Squamous Cell Carcinomas in Uighur Women

  • Kurban, Shajidai;Tursun, Mikrban;Kurban, Gulinar;Hasim, Ayshamgul
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권21호
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    • pp.9211-9216
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    • 2014
  • CXCR7 is involved in tumor development and metastasis in multiple malignancies. However, the function and molecular mechanisms of action of CXCR7 in human cervical cancer are still unclear. In the present study a loss of-function approach was used to observe the effects of recombinant CXCR7 specific small interfering RNA pBSilence1.1 plasmids on biological behavior including proliferative activity and invasive potential, as indicated by MTT assays with the cervical cancer SiHa cell line in vitro. Reverse transcription polymerase chain reaction and Western blotting revealed that CXCR7 was downregulated in transfected compared with control cells, associated with inhibited cell growth, invasiveness and migration. The expression of CXCR7 and CXCL12 was also determined immunohistochemically in 152 paraffin-embedded, cervical squamous cell carcinoma (CSCC) and cervical intraepithelial neoplasia (CIN), or normal cervical epithelial to assess clinico-pathological pattern and CXCR7 status with respect to cell differentiation and lymph node metastasis in Uighur patients with CSCC. CXCR7 and CXCL12 expression was higher in cervical cancer than CIN and normal cervical mucosa, especially in those with higher stage and lymph node metastasis. CXCL12 appeared to be positively regulated by CXCR7 at the post-transcriptional level in CSCC. We propose that aberrant expression of CXCR7 plays a role in carcinogenesis, differentiation and metastasis of CSCC, implying its use as a potential target for clinical biomarkers in differentiation and lymph node metastasis.

Characteristics of Purinergic Receptor Expressed in 3T3-L1 Preadipocytes

  • ;;;;공인덕
    • 대한의생명과학회지
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    • 제15권4호
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    • pp.319-326
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    • 2009
  • Extracellular ATP elicits diverse physiological effects by binding to the G-protein-coupled P2Y receptors on the plasma membrane. In addition to the short-term effects of extracellular nucleotides on cell functions, there is evidence that such purinergic signalling can have long-term effects on cell proliferation, differentiation and death. The 3T3-L1 cell line derived from mouse embryo is a well-established and commonly utilized in vitro model for adipocytes differentiation and function. However, the distributions and roles of P2Y subtypes are still unknown in the preadipocyte. In this study, we identified the distributions and roles of P2Y subtypes in preadipocyte using $Ca^{2+}$ imaging and realtime PCR. ATP increased the $[Ca^{2+}]_i$ in a concentration-dependent manner. ATP increased $Ca^{2+}$ in absence and/or presence of extracellular $Ca^{2+}$. Suramin, non-selective P2Y blocker, largely blocked the ATP-induced $Ca^{2+}$ response. U73122, a PLC inhibitor, completely inhibited $Ca^{2+}$ mobilization in 3T3-L1 cells. The mRNA expression by realtime PCR of P2Y subtypes was $P2Y_2:P2Y_5:P2Y_6=1.0:12.5:0.3$. In conclusion, we showed that $P2Y_5$ receptor is a dominant purinergic receptor in preadipocytes, and multiple P2Y receptors could involve in differentiation and migration via regulating of intracellular calcium concentration.

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Swarming Differentiation of Vibrio vulnificus Downregulates the Expression of the vvhBA Hemolysin Gene via the LuxS Quorum-Sensing System

  • Kim Moon-Young;Park Ra-Young;Choi Mi-Hwa;Sun Hui-Yu;Kim Choon-Mee;Kim Soo-Young;Rhee Joon-Haeng;Shin Sung-Heui
    • Journal of Microbiology
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    • 제44권2호
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    • pp.226-232
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    • 2006
  • Swarming has proven to be a good in vitro model for bacterial surface adherence and colonization, and the swarming differentiation of a bacterium has been shown to be coupled with changes in the expression of virulence factors associated with its invasiveness, particularly in the early stages of infection. In this study, we attempted to determine whether the expression of vvhA, which encodes for hemolysin/cytolysin (VvhA), is either upregulated or downregulated during the swarming differentiation of V. vulnificus. The insertional inactivation of vvhA itself exerted no detectable effect on the expression of V. vulnificus swarming motility. However, in our lacZ-fused vvhA transcriptional reporter assay, vvhA expression decreased in swarming V. vulnificus as compared to non-swarming or planktonic V. vulnificus. The reduced expression of vvhA in swarming V. vulnificus increased as a result of the deletional inactivation of luxS, a gene associated with quorum sensing. These results show that vvhA expression in swarming V. vulnificus is downregulated via the activity of the LuxS quorum-sensing system, suggesting that VvhA performs no essential role in the invasiveness of V. vulnificus via the adherence to and colonization on the body surfaces required in the early stages of the infection. However, VvhA may playa significant role in the pathophysiological deterioration occurring after swarming V. vulnificus is differentiated into planktonic V. vulnificus.

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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    • 제8권1호
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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.

블라스팅과 양극산화된 티타늄 표면이 조골세포의 부착 및 분화에 미치는 영향 (Effect of Blasted or Anodized Titanium Surface Roughness on Adhesion and Differentiation of Osteoblasts)

  • 박찬진;조리라;이양진;고성희
    • 구강회복응용과학지
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    • 제22권3호
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    • pp.261-270
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    • 2006
  • The success of an implant is determined by its integration into the tissue surrounding the biomaterial. Surface roughness is considered to influence the behavior of adherent cells. The aim of this in vitro study was to determine the effect of surface roughness on Saos-2 osteoblast-like cells. Titanium disks, blasted with $75{\mu}m$ aluminum oxide particles and anodic oxidized and machined titanium disks were prepared. Saos-2 were plated on the disks at a density of 50,000 cells per well in 48-well dishes. After 1 hour, 1 day, 6 days cell numbers were counted. One day, 6 days after plating, alkaline phosphatase(ALPase) activity was determined. Compared to experimental groups, the number of cells was significantly higher on control group. The stimulatory effect of surface roughness on ALPase was more pronounced on the experimental groups than on control group. These results demonstrate that surface roughness alters proliferation and differentiation of osteoblasts. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells.

티타늄 표면조도가 조골세포의 부착 및 분화에 미치는 영향 (Effect of titanium surface roughness on adhesion and differentiation of osteoblasts)

  • 김정식;이재관;고성희;엄흥식;장범석
    • Journal of Periodontal and Implant Science
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    • 제35권4호
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    • pp.839-850
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    • 2005
  • The success of an implant is determined by its integration into the tissue surrounding the biomaterial. Surface roughness is considered to influence the behavior of adherent cells. The aim of this in vitro study was to determine the effect of surface roughness on Saos-2 osteoblast-like cells. Titanium disks blasted with 75 ${/mu}m$ aluminum oxide particles and machined titanium disks were prepared. Saos-2 were plated on the disks at a density of 50,000 cells per well in 48-well dishes. After 1 hour, 1 day, 6 days cell numbers were counted. One day, 6 days after plating, alkaline phosphatase(ALPase) activity was determined. Compared to experimental group, the number of cells was significantly higher on control group. The stimulatory effect of surface roughness on ALPase was more pronounced on the experimental group than on control group. These results demonstrate that surface roughness alters proliferation and differentiation of osteoblasts. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells.

Inhibitory effect of Ssanghwa-tang on bone loss in ovariectomized rats

  • Shim, Ki-Shuk;Lee, Ji-Hye;Ma, Choong-Je;Lee, Yoon-Hee;Choi, Sung-Up;Lee, Jae-Hoon;Ma, Jin-Yeul
    • Animal cells and systems
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    • 제14권4호
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    • pp.283-289
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    • 2010
  • Ssanghwa-tang (SHT) is a traditional Korean herbal medicine widely prescribed to decrease fatigue following an illness. The purpose of this study was to investigate the effects of SHT on osteoclast differentiation in vitro, and on bone loss in ovariectomized (OVX) rats in vivo. SHT significantly reduced the receptor activator for the nuclear factor ${\kappa}B$ (NF-${\kappa}B$) ligand (RANKL)-induced tartrate-resistant acid phosphatase (TRAP) activity, and multinucleated osteoclast formation in RAW264.7 cells without affecting cell viability. In addition, SHT significantly attenuated RANKL-induced mRNA expression levels of c-Src and cathepsin K. To examine the in vivo effect of SHT on OVX-induced bone loss in OVX rats, we administered SHT (0.6 g/kg BID) orally to OVX rats for 12 weeks. SHT administration significantly blocked OVX-induced decrease of femoral bone mineral density (BMD) and femoral trabeculae in OVX rats. In conclusion, these results suggest that SHT treatment effectively prevents OVX-induced bone loss, and this effect may result from its inhibitory effect on osteoclast differentiation.