The Experts below are selected from a list of 61602 Experts worldwide ranked by ideXlab platform
Moonsik Yang - One of the best experts on this subject based on the ideXlab platform.
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co expression of proteinase inhibitor enhances recombinant human granulocyte macrophage colony stimulating factor production in transgenic rice Cell Suspension culture
Protein Expression and Purification, 2008Co-Authors: Taegeum Kim, Taeho Kwon, Yongsuk Jang, Hojin Lee, Yunji Shin, Moonsik YangAbstract:Abstract The synthetic gene (sPI-II) harboring the chymotrypsin (C1) and trypsin (T1) inhibitor domains of the Nicotiana alata serine proteinase inhibitor II gene has been previously expressed, and extraCellular protease activity was shown to be reduced in the Suspension culture medium. In this study, the sPI-II gene was introduced into transgenic rice Cells expressing rhGM-CSF (recombinant human granulocyte–macrophage colony-stimulating factor), in an effort to reduce protease activity and increase rhGM-CSF accumulation in the Suspension culture medium. The integration and expression of the introduced sPI-II gene in the transgenic rice Cells were verified via genomic DNA PCR amplification and Northern blot analysis, respectively. Relative protease activity was found to have been reduced and rhGM-CSF production was increased 2-fold in the co-transformed Cell Suspension culture with rhGM-CSF and the sPI-II gene, as compared with that observed in the transformed Cell Suspension culture expressing rhGM-CSF only. These results indicate that a transformed plant Cell Suspension culture system expressing the proteinase inhibitor can be a useful tool for increasing recombinant protein production.
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production of bioactive human granulocyte colony stimulating factor in transgenic rice Cell Suspension cultures
Protein Expression and Purification, 2006Co-Authors: Shinyoung Hong, Taeho Kwon, Yongsuk Jang, Sooho Kim, Moonsik YangAbstract:Human granulocyte-colony stimulating factor (hG-CSF), a human cytokine, was expressed in transgenic rice Cell Suspension culture. The hG-CSF gene was cloned into the rice expression vector containing the promoter, signal peptide, and terminator derived from a rice α-amylase gene Amy3D. Using particle bombardment-mediated transformation, hG-CSF gene was introduced into the calli of rice (Oryza sativa) cultivar Dong-jin. Expression of the hG-CSF gene was confirmed by ELISA and Northern blot analysis. The amount of recombinant hG-CSF accumulated in culture medium from transgenic rice Cell Suspension culture on the sugar starvation was determined by time series ELISA. Biological activity of the plant derived hG-CSF was confirmed by measuring the proliferation of the AML-193 Cells, and was similar to that of the commercial Escherichia coli-derived hG-CSF. In this paper, we discuss the attractive attributes of using rice Cell Suspension system for the expression of therapeutic recombinant hG-CSF.
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high level of expression of recombinant human granulocyte macrophage colony stimulating factor in transgenic rice Cell Suspension culture
Biotechnology and Bioengineering, 2003Co-Authors: Yunji Shin, Shinyoung Hong, Taeho Kwon, Yongsuk Jang, Moonsik YangAbstract:Recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) has been previously produced in tobacco Cell Suspension cultures. However, the amount of hGM-CSF accumulated in the culture medium dropped quickly from its maximum of 150 microg/L at 5 d after incubation. To overcome this problem, we sought an expression system in which heterologous gene expression could be induced at high levels. We selected a rice amylase expression system in which the promoter Ramy3D is induced to express recombinant protein by sucrose starvation. This induction system was found to give good yield of recombinant hGM-CSF in transgenic rice Cell Suspension culture and protease activity of this culture medium was low compared to that of tobacco culture system.
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establishment of a transgenic tobacco Cell Suspension culture system for producing murine granulocyte macrophage colony stimulating factor
Molecules and Cells, 1997Co-Authors: Joonggi Lee, Taeho Kwon, Sangbong Choi, H S Kang, Kangjin Cho, Dae Hyuk Kim, Youngjoo Jang, Moonsik YangAbstract:We tested if murine granulocyte-macrophage colony stimulating factor (mGM-CSF) is produced as a biologically active form through plant Cell culture. The mGM-CSF gene was cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring the recombinant mGM-CSF (rmGM-CSF) gene. Cell Suspension culture was established from the leaf-derived calli of transgenic tobacco plants. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and Cell Suspension cultures. In addition, the biological activity of rmGM-CSF from plant Cell culture was confirmed by measuring the proliferation of GM-CSF dependent FDC-P1 Cells.
Somesh Gupta - One of the best experts on this subject based on the ideXlab platform.
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extracted hair follicle outer root sheath Cell Suspension for pigment Cell restoration in vitiligo
Journal of Cutaneous and Aesthetic Surgery, 2013Co-Authors: Anil Kumar, Sujata Mohanty, Kanika Sahni, Rajesh Kumar, Somesh GuptaAbstract:Vitiligo surgery has come up a long way from punch skin grafts to epidermal Cell Suspension and latest to the extracted hair follicle outer root sheath Cell Suspension (EHF-ORS-CS) transplantation. The progressive development from one technique to the other is always in a quest for the best. In the latest development- EHF-ORS-CS, which is an enriched source of follicular inactive melanocyte (melanocyte stem Cells), seems to be a good addition to the prevailing Cell-based therapies for vitiligo; however, need to be explored further in larger, and preferably randomized blinded studies. This review discusses the principle, technical details, and stem Cell composition of hair follicular outer root sheath Cell Suspension.
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noncultured extracted hair follicle outer root sheath Cell Suspension for transplantation in vitiligo
British Journal of Dermatology, 2011Co-Authors: Sujata Mohanty, Anil Kumar, Jyoti Dhawan, V Sreenivas, Somesh GuptaAbstract:Summary Background Current noncultured Cell-based transplantation therapies for vitiligo largely involve shave skin biopsy for preparation of noncultured melanocyte Suspension. As the overall proportion of melanocytes is low in the epidermis, these techniques require basal Cell layer enrichment, which adds additional steps. We tried follicular unit extraction (FUE) to harvest hair follicles as a source of melanocytes. Objectives To evaluate the efficacy of a novel surgical method for vitiligo: noncultured extracted hair follicular outer root sheath (ORS) Cell Suspension transplantation. Methods Fourteen patients with vitiligo, stable for at least 3 months, were included in this prospective study. Fifteen to 25 hair follicles were extracted from occipital scalp using the FUE method. Hair follicles were incubated with trypsin–ethylenediamine tetraacetic acid solution at 37 °C for 90 min to separate ORS Cells. The Cell Suspension was filtered through a 70-μm Cell strainer, then centrifuged for 5 min at 1000 r.p.m. to obtain a Cell pellet. The pellet was resuspended and applied to the dermabraded recipient area and dressed. Results The mean ± SD repigmentation was 65·7 ± 36·7%. Overall, nine of 14 patients achieved > 75% repigmentation. Mean percentage repigmentation was significantly higher in patients with ≥ 1 year stability than those with < 1 year stability (P = 0·02). Conclusions Extracted hair follicular ORS Cell Suspension can be a useful simplified transplantation method for vitiligo. The transplantation procedure should be reserved for patients with vitiligo stable for at least 1 year. A larger study is needed for further evaluation.
Sujata Mohanty - One of the best experts on this subject based on the ideXlab platform.
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extracted hair follicle outer root sheath Cell Suspension for pigment Cell restoration in vitiligo
Journal of Cutaneous and Aesthetic Surgery, 2013Co-Authors: Anil Kumar, Sujata Mohanty, Kanika Sahni, Rajesh Kumar, Somesh GuptaAbstract:Vitiligo surgery has come up a long way from punch skin grafts to epidermal Cell Suspension and latest to the extracted hair follicle outer root sheath Cell Suspension (EHF-ORS-CS) transplantation. The progressive development from one technique to the other is always in a quest for the best. In the latest development- EHF-ORS-CS, which is an enriched source of follicular inactive melanocyte (melanocyte stem Cells), seems to be a good addition to the prevailing Cell-based therapies for vitiligo; however, need to be explored further in larger, and preferably randomized blinded studies. This review discusses the principle, technical details, and stem Cell composition of hair follicular outer root sheath Cell Suspension.
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noncultured extracted hair follicle outer root sheath Cell Suspension for transplantation in vitiligo
British Journal of Dermatology, 2011Co-Authors: Sujata Mohanty, Anil Kumar, Jyoti Dhawan, V Sreenivas, Somesh GuptaAbstract:Summary Background Current noncultured Cell-based transplantation therapies for vitiligo largely involve shave skin biopsy for preparation of noncultured melanocyte Suspension. As the overall proportion of melanocytes is low in the epidermis, these techniques require basal Cell layer enrichment, which adds additional steps. We tried follicular unit extraction (FUE) to harvest hair follicles as a source of melanocytes. Objectives To evaluate the efficacy of a novel surgical method for vitiligo: noncultured extracted hair follicular outer root sheath (ORS) Cell Suspension transplantation. Methods Fourteen patients with vitiligo, stable for at least 3 months, were included in this prospective study. Fifteen to 25 hair follicles were extracted from occipital scalp using the FUE method. Hair follicles were incubated with trypsin–ethylenediamine tetraacetic acid solution at 37 °C for 90 min to separate ORS Cells. The Cell Suspension was filtered through a 70-μm Cell strainer, then centrifuged for 5 min at 1000 r.p.m. to obtain a Cell pellet. The pellet was resuspended and applied to the dermabraded recipient area and dressed. Results The mean ± SD repigmentation was 65·7 ± 36·7%. Overall, nine of 14 patients achieved > 75% repigmentation. Mean percentage repigmentation was significantly higher in patients with ≥ 1 year stability than those with < 1 year stability (P = 0·02). Conclusions Extracted hair follicular ORS Cell Suspension can be a useful simplified transplantation method for vitiligo. The transplantation procedure should be reserved for patients with vitiligo stable for at least 1 year. A larger study is needed for further evaluation.
Taeho Kwon - One of the best experts on this subject based on the ideXlab platform.
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co expression of proteinase inhibitor enhances recombinant human granulocyte macrophage colony stimulating factor production in transgenic rice Cell Suspension culture
Protein Expression and Purification, 2008Co-Authors: Taegeum Kim, Taeho Kwon, Yongsuk Jang, Hojin Lee, Yunji Shin, Moonsik YangAbstract:Abstract The synthetic gene (sPI-II) harboring the chymotrypsin (C1) and trypsin (T1) inhibitor domains of the Nicotiana alata serine proteinase inhibitor II gene has been previously expressed, and extraCellular protease activity was shown to be reduced in the Suspension culture medium. In this study, the sPI-II gene was introduced into transgenic rice Cells expressing rhGM-CSF (recombinant human granulocyte–macrophage colony-stimulating factor), in an effort to reduce protease activity and increase rhGM-CSF accumulation in the Suspension culture medium. The integration and expression of the introduced sPI-II gene in the transgenic rice Cells were verified via genomic DNA PCR amplification and Northern blot analysis, respectively. Relative protease activity was found to have been reduced and rhGM-CSF production was increased 2-fold in the co-transformed Cell Suspension culture with rhGM-CSF and the sPI-II gene, as compared with that observed in the transformed Cell Suspension culture expressing rhGM-CSF only. These results indicate that a transformed plant Cell Suspension culture system expressing the proteinase inhibitor can be a useful tool for increasing recombinant protein production.
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production of bioactive human granulocyte colony stimulating factor in transgenic rice Cell Suspension cultures
Protein Expression and Purification, 2006Co-Authors: Shinyoung Hong, Taeho Kwon, Yongsuk Jang, Sooho Kim, Moonsik YangAbstract:Human granulocyte-colony stimulating factor (hG-CSF), a human cytokine, was expressed in transgenic rice Cell Suspension culture. The hG-CSF gene was cloned into the rice expression vector containing the promoter, signal peptide, and terminator derived from a rice α-amylase gene Amy3D. Using particle bombardment-mediated transformation, hG-CSF gene was introduced into the calli of rice (Oryza sativa) cultivar Dong-jin. Expression of the hG-CSF gene was confirmed by ELISA and Northern blot analysis. The amount of recombinant hG-CSF accumulated in culture medium from transgenic rice Cell Suspension culture on the sugar starvation was determined by time series ELISA. Biological activity of the plant derived hG-CSF was confirmed by measuring the proliferation of the AML-193 Cells, and was similar to that of the commercial Escherichia coli-derived hG-CSF. In this paper, we discuss the attractive attributes of using rice Cell Suspension system for the expression of therapeutic recombinant hG-CSF.
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high level of expression of recombinant human granulocyte macrophage colony stimulating factor in transgenic rice Cell Suspension culture
Biotechnology and Bioengineering, 2003Co-Authors: Yunji Shin, Shinyoung Hong, Taeho Kwon, Yongsuk Jang, Moonsik YangAbstract:Recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) has been previously produced in tobacco Cell Suspension cultures. However, the amount of hGM-CSF accumulated in the culture medium dropped quickly from its maximum of 150 microg/L at 5 d after incubation. To overcome this problem, we sought an expression system in which heterologous gene expression could be induced at high levels. We selected a rice amylase expression system in which the promoter Ramy3D is induced to express recombinant protein by sucrose starvation. This induction system was found to give good yield of recombinant hGM-CSF in transgenic rice Cell Suspension culture and protease activity of this culture medium was low compared to that of tobacco culture system.
-
establishment of a transgenic tobacco Cell Suspension culture system for producing murine granulocyte macrophage colony stimulating factor
Molecules and Cells, 1997Co-Authors: Joonggi Lee, Taeho Kwon, Sangbong Choi, H S Kang, Kangjin Cho, Dae Hyuk Kim, Youngjoo Jang, Moonsik YangAbstract:We tested if murine granulocyte-macrophage colony stimulating factor (mGM-CSF) is produced as a biologically active form through plant Cell culture. The mGM-CSF gene was cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring the recombinant mGM-CSF (rmGM-CSF) gene. Cell Suspension culture was established from the leaf-derived calli of transgenic tobacco plants. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and Cell Suspension cultures. In addition, the biological activity of rmGM-CSF from plant Cell culture was confirmed by measuring the proliferation of GM-CSF dependent FDC-P1 Cells.
Anil Kumar - One of the best experts on this subject based on the ideXlab platform.
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extracted hair follicle outer root sheath Cell Suspension for pigment Cell restoration in vitiligo
Journal of Cutaneous and Aesthetic Surgery, 2013Co-Authors: Anil Kumar, Sujata Mohanty, Kanika Sahni, Rajesh Kumar, Somesh GuptaAbstract:Vitiligo surgery has come up a long way from punch skin grafts to epidermal Cell Suspension and latest to the extracted hair follicle outer root sheath Cell Suspension (EHF-ORS-CS) transplantation. The progressive development from one technique to the other is always in a quest for the best. In the latest development- EHF-ORS-CS, which is an enriched source of follicular inactive melanocyte (melanocyte stem Cells), seems to be a good addition to the prevailing Cell-based therapies for vitiligo; however, need to be explored further in larger, and preferably randomized blinded studies. This review discusses the principle, technical details, and stem Cell composition of hair follicular outer root sheath Cell Suspension.
-
noncultured extracted hair follicle outer root sheath Cell Suspension for transplantation in vitiligo
British Journal of Dermatology, 2011Co-Authors: Sujata Mohanty, Anil Kumar, Jyoti Dhawan, V Sreenivas, Somesh GuptaAbstract:Summary Background Current noncultured Cell-based transplantation therapies for vitiligo largely involve shave skin biopsy for preparation of noncultured melanocyte Suspension. As the overall proportion of melanocytes is low in the epidermis, these techniques require basal Cell layer enrichment, which adds additional steps. We tried follicular unit extraction (FUE) to harvest hair follicles as a source of melanocytes. Objectives To evaluate the efficacy of a novel surgical method for vitiligo: noncultured extracted hair follicular outer root sheath (ORS) Cell Suspension transplantation. Methods Fourteen patients with vitiligo, stable for at least 3 months, were included in this prospective study. Fifteen to 25 hair follicles were extracted from occipital scalp using the FUE method. Hair follicles were incubated with trypsin–ethylenediamine tetraacetic acid solution at 37 °C for 90 min to separate ORS Cells. The Cell Suspension was filtered through a 70-μm Cell strainer, then centrifuged for 5 min at 1000 r.p.m. to obtain a Cell pellet. The pellet was resuspended and applied to the dermabraded recipient area and dressed. Results The mean ± SD repigmentation was 65·7 ± 36·7%. Overall, nine of 14 patients achieved > 75% repigmentation. Mean percentage repigmentation was significantly higher in patients with ≥ 1 year stability than those with < 1 year stability (P = 0·02). Conclusions Extracted hair follicular ORS Cell Suspension can be a useful simplified transplantation method for vitiligo. The transplantation procedure should be reserved for patients with vitiligo stable for at least 1 year. A larger study is needed for further evaluation.