• Journal of Animal Science and Technology
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• 제46권2호
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• pp.155-164
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• 2004

이 연구는 gene-trap construct를 가지고 있는 배양세포로부터 trap gene을 확인하고 클로닝한 다음 이러한 세포를 이용하여 복제 지브라물고기를 만들기 위해 수행되어졌다. 본 연구에서 gene-trap과 연관된 복제 지브라물고기가 성공적으로 만들어졌다. 본 실험에서 두 종류의 백터(SA/GFP-TP와 Neo-TP)가 사용되었다. 이들 벡터에 의해 전이된 모든 종류의 세포는 항생제에 의해 선별을 하여 분석에 이용하였다. SA/GFP-TP에 의해 전이된 세포의 경우, 단일세포상에서 GFP 발현도가 낮아 본 연구에서 동물복제에 사용되지 않았으며, Neo-TP에 의해 전이된 세포주가 복제실험에 이용되었다. Neo-TP 세포에 의한 복제실험 결과, 총 1179개의 핵치환 난으로부터 44(3.7%) 개의 배자가 포배기에 도달하였으며, 8(0.8%) 개의 배자가 부화시기에 이르렀다. 그리고 3마리는 성숙단계에 이르렀으며, 이중 1마리에서 정상적으로 gene-trap 전이가 이루어짐을 Southern blot 분석을 통해 확인되었다.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2000년도 제28회 학술심포지엄
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• pp.11-14
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• 2000

• 대한예방한의학회지
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• 제14권3호
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• pp.13-26
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• 2010

Objectives : This study was performed to evaluate the effect of Securinega suffructicosa Rehder (SS) on osteoclast differentiation and gene expression. Methods : The osteoclastogenesis and gene expression were determined in RANKL-stimulated RAW 264.7. The results were summarized as followes. Results : SS decreased the number of TRAP positive cell in RANKL-stimulated RAW264.7 cell. SS decreased the expression of RANK, TNF${\alpha}$, and IL-6 in RANKL-stimulated RAW264.7 cell. SS decreased the expression of iNOS and COX-2 in RANKL-stimulated RAW264.7 cell. SS decreased the expression of Cathepsin K in RANKL-stimulated RAW264.7 cell. Conclusions : It is concluded that SS might decrease the bone resorption resulted from decrease of osteoclast differentiation and it's related gene expression.

• BMB Reports
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• 제43권12호
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• pp.789-794
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• 2010

A novel gene, designated mgt-6, containing four splicing variants, was isolated from a gene trap clone library of C3H/10T1/2 cells transfected with retroviral promoterless gene-trap vector, ROSAFARY. The transcript variants were differentially expressed in murine tissues and cell lines and differentially responded to diverse stimuli including TGF-${\beta}1$ and mitogen-activated protein kinase (MAPK) inhibitors. The mgt-6 gene encoded a protein of 37 or 11 amino acid residuals with cytoplasmic distribution. However, when C3H/10T1/2 cells were treated with 5-azacytidine, the protein translocated into cell nucleus as indicated by fused LacZ or C-terminally tagged EGFP. Our preliminary results suggest that further study on the role of mgt-6 gene in cell transformation and differentiation may be of significance.

• 대한한방부인과학회지
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• 제25권3호
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• pp.16-26
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• 2012

Objectives: This study was performed to evaluate effects of Cordyceps militaris(CM) on osteoclast differentiation and its related gene expression. Methods: We used mouse myeloid cells RAW 264.7 stimulated by receptor activator of nuclear factor kappa-B ligand(RANKL) to induce osteoclast differentiation. There are four groups of which RAW 264.7 cells are not stimulated by RANKL (Normal), stimulated by RANKL without CM(Control), stimulated by RANKL with 0.1 ${\mu}g/ml$ of CM(CM 0.1), stimulated by RANKL with 1 ${\mu}g/ml$ of CM(CM 1). Osteoclastogenesis was measured by counting Tartrate-resistant acid phosphatase-positive multinucleated cells [TRAP(+) MNC]. RT-PCR was performed to evaluate the inhibitory effect of CM on gene expression(TRAP, AKT1, JNK1, NFATc1, c-Fos, MITF). Results: 1. CM decreased the number of TRAP(+) osteoclast in RANKL-stimulated RAW 264.7 cell at the concentration of 0.1 ${\mu}g/ml$ and 1 ${\mu}g/ml$. 2. CM decreased the expression of TRAP in osteoclast at the concentration of 1 ${\mu}g/ml$. 3. CM decreased the expression of AKT1, JNK1 in osteoclast at the concentration of 1 ${\mu}g/ml$. 4. CM didn't affect the expression of NFATc1, c-Fos, MITF in osteoclast. Conclusions: Cordyceps militaris has inhibitory effects on osteoclast differentiation and its related gene expression. These results suggest that Cordyceps militaris has a potential as a treatment of osteoporosis.

• 대한한방부인과학회지
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• 제25권2호
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• pp.23-42
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• 2012

Objectives: This study was performed to evaluate the effect of Samkieumgamibang (SKG) on osteoporosis. Methods: The osteoclastogenesis and gene expression were determined in RANKL-stimulated RAW 264.7 cell. And, osteoblastogenesis was also determined in rat calvarial cell. Results: SKG decreased the number of TRAP positive cell in osteoclast. It also decreased the expression of Cathepsin K, MMP-9, TRAP, c-fos, NAFTc1 and JNK1 in osteoclast. SKG increased the expression of iNOS in RANKL-stimulated in osteoclast. Otherwise, SKG inhibited TRAP activity in osteoclast. SKG increased cell proliferation, ALP activity, bone martix protein, collagen and nodule in osteoblast. Conclusions: It is concluded that SKG might decrease the bone resorption resulted from decrease of osteoclast differentiation and it's related gene expression. And, SKG might increase the bone formation resulted from increase of osteoblast function.

• 대한한방내과학회지
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• 제38권6호
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• pp.1035-1048
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• 2017

Objectives: This study was performed to investigate the effects of Gardenia jasminoides extract (GJ) on osteoclast differentiation and bone resorption in vitro. Methods: To investigate the effect of GJ on osteoclast differentiation, the mouse leukemic myeloid cell line RAW 264.7 was stimulated by RANKL (receptor activator of nuclear factor kB ligand). Osteoclast differentiation was measured by counting TRAP (+) MNC in the presence of RANKL. To elucidate the mechanism of the inhibitory effect of GJ on osteoclast differentiation, gene expression of TRAP, Cathepsin K, MMP-9, NFATc1, c-Fos, MITF, DC-STAMP, CTR, OC-STAMP and Atp6v0d2 was measured using reverse transcription-PCR (RT-PCR). Bone resorption was measured using the bone pit formation assay. Results: GJ decreased the number of TRAP (+) MNCs in the presence of RANKL. GJ inhibited the expression of cathepsin K, MMP-9, TRAP, MITF, NFATc1, c-Fos, iNON, OC-STAMP, Atp6v0d2, and DC-STAMP in the osteoclast, and inhibited bone pit formation in vitro. Conclusions: The results suggest that GJ has inhibitory effects on bone resorption resulting from inhibition of osteoclast differentiation and gene expression.

• 대한한방부인과학회지
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• 제23권3호
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• pp.78-90
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• 2010

Purpose: This study was performed to evaluate the effect of Clematidis Radix extract(CB) on osteoclast differentiation and gene expression. The osteocastogenesis and gene expression were determined in RANKL-induced RAW 264.7 cell. Methods: RANKL-induced RAW 264.7 cell with Clematidis Radix extract was stained by TRAP which is expressive marker of osteoclast. The gene expression of RANK, $TNF{\alpha}$, IL-6, iNOS and Cathepsin, those are factors related to bone resorption, was estimated by using Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Results: Clematidis Radix extract decreased the number of TRAP-positive multi nuclei cell, and decreased the gene expression of RANK, $TNF{\alpha}$, IL-6, iNOS and Cathepsin K in RANKL-induced RAW 264.7 cell. Conclusion: It is concluded that Clematidis Radix extract might decrease the bone resorption resulted from decrease of osteoclast differentiation and it's related gene expression.

• 대한한방부인과학회지
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• 제27권2호
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• pp.59-70
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• 2014

Purpose: This study was conducted to evaluate the inhibitory effect of ursolic acid from Prunella vulgaris on osteoclast differentiation. Methods: MTT-assay was performed to estimate cytotoxicity of ursolic acid from Prunella vulgaris in BMMs stimulated with M-CSF. TRAP staining, TRAP activity and Real-time PCR were performed to know the inhibitory effect on osteoclast differentiation. Actin ring formation were analysed to observe the effect of ursolic acid from Prunella vulgaris. Results: Ursolic acid from Prunella vulgaris has no cytotoxicity at the concentration of $1{\mu}g/ml$ or lower. Ursolic acid decreased the number of TRAP positive cells and the expression of NFATc1 gene, c-Fos gene, TRAP and OSCAR in BMMs stimulated with RANKL. Ursolic acid restrained the formation of actin ring. Ursolic acid inhibited NF-${\kappa}B$ activity by inducing degradation of p-$IkB{\alpha}$. Conclusions: Ursolic acid from Prunella vulgaris has the inhibitory effect of osteoclast differentiation and bone resorption. Futher studies are needed to treat osteoporosis by usolic acid from Prunella vulgaris.

• 대한한방부인과학회지
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• 제25권2호
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• pp.1-11
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• 2012

Objectives: This study was conducted to evaluate the inhibitory effect of Melia Fructus extract on osteoclast differentiation. Methods: MTT-assay was performed to estimate cytotoxicity of Melia Fructus extract in BMMs stimulated with M-CSF. TRAP staining, TRAP activity and Real-time PCR were performed to know the inhibitory effect on osteoclast differentiation. Actin ring formation were analysed to observe the effect of Melia Fructus extract. Results: Melia Fructus extract decreased the number of TRAP positive cells and the expression of NFATc1 gene, c-Fos gene, TRAP and OSCAR in BMMs stimulated with RANKL. Melia Fructus extract has no cytotoxicity at the concentration used in this study. Melia Fructus extract restrained the formation of actin ring. Melia Fructus inhibited NF-${\kappa}B$ activity by inducing degradation of p-$IkB{\alpha}$. Conclusions: Melia Fructus has the inhibitory effect of osteocalst differentiation and bone resorption. Further studies are needed to treat osteoporosis by herbal medicine containing Melia Fructus.