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Study the immunomodulatory activity of Semen cuscutae and identify the active components
Abstracts / Journal of Bioscience and Bioengineering 108 (2009) S4–S20 free media, although inferior to one of the media containing mammalderived factors. Furthermore, Sericin-GIT medium was tested to culture various cell lines such as HepG2, HeLa, SIRC and L929 cell lines and compared with two serum-free media and one serum-containing medium. HepG2 cells cultured in Sericin-GIT medium proliferated better than those in other media, and other three cell lines in SericinGIT medium proliferated as well as in other medium. These results indicate that Sericin-GIT medium is suitable for the cell culture for regenerative medicine and protein pharmaceutical production. doi:10.1016/j.jbiosc.2009.08.490
AN-P34 The generation of oligodendrocytes from induced pluripotent stem cells Shinichiro Ogawa,1 Yasuhito Tokumoto,1 Jun Miyake,1,2 and Teruyuki Nagamune1
Oligodendrocytes are one type of neuronal glia cells in the central nervous system and form myelin sheaths insulating neuronal axons. Oligodendrocytes are such important cells, so the method of generating these cells has been developed for treating demyelinating disease. Recently, induced pluripotent stem (iPS) cells are generated. Although some types of cells are reported to be generated from iPS cells, the success of being differentiated into oligodendrocytes has not been reported yet. So, we tested whether iPS cells could be differentiated into oligodendrocytes by the differentiation method for ES cells and whether there were some differences in the differentiation efficiency between iPS cells and ES cells. As a result, we succeeded in generating oligodendrocytes from iPS cells. However, the differentiation efficiency of iPS cells was 2% and lower than that of ES cells. This result indicates that some factors of iPS cells inhibit differentiation and that iPS cells are not the same as ES cells. Reference 1. Yasuhito Tokumoto, Shinichiro Ogawa, Teruyuki Nagamune, Jun Miyake: “A comparison of efficiency of in vitro differentiation of oligodendrocytes from induced pluripotent stem cells versus embryonic stem cells” (in revision Experimental Cell Research, 2009).
doi:10.1016/j.jbiosc.2009.08.491
AN-P35 Study the immunomodulatory activity of Semen cuscutae and identify the active components 1
Ming-Kuem Lin, Meng-Shiou Lee, Yang,1 and Ching-Liang Chu2
Cuscuta seed, also known as Semen cuscutae or Tu-Si-Zi, is one of the commonly used Chinese medicinal materials in Asia. It is mostly used for kidney deficiency. However, the immunomodulatory activity of S. cuscutae is not known. Since dendritic cells (DCs) play a key role in regulating immune responses, we examined the effect of S. cuscutae on DCs. Mouse bone marrow-derived DCs were treated with the methanol extract of S. cuscutae (MESC), and we found that MESC induced the pro-inflammatory cytokine production by DCs, suggesting that MESC can potentially activate DCs. In addition, MESC promoted DC maturation by enhancing the expression of maturation markers. To identify the active ingredient, we further purified several compounds from MESC and tested their effect on DC activation. Unfortunately, we did not find any active candidate in first screen yet. Remarkably, our study demonstrates the immunomodulatory activity of S. cuscutae for the first time and then extends its biological function. It is interesting to reveal the molecular mechanism for this DC activation by finding the active component for further application in pharmaceutical industry. doi:10.1016/j.jbiosc.2009.08.492
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan 1 and Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka University, Osaka 560-8531, Japan 2
1
S19
1
Wen-Te Chang,
Meng-Chia
School of Chinese Medicine Resources, China Medical University, Taichung, Taiwan 1 and Immunology Research Center, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan 2
AN-P37 The production of swine cholera marker vaccine by serum free culture of insect cells infected with baculovirus Jeong Nam Park, Je Sin Woo, Chang Jin Lee, and Yeon Ho Jeong Kangwon National University, Chuncheon, Republic of Korea The development of porcine cholera marker vaccine is needed for the maintenance of porcine cholera free region. Insect cells are used as host cells for the expression of marker vaccine by baculovirus vector system. Insect cell lines are able to grow suspension culture and easy to scale up. The use of FBS has several disadvantages and is undesirable in large scale insect cell cultures media. Therefore, high density insect cell culture in serum free medium is prerequisite to produce a marker vaccine with high productivity and safety. Therefore, it is important to select the optimum serum free medium for host cell line. In this work, two insect cell lines, Hi5 originated from the eggs of Tricoplusiu ni and Sf9 derived from the ovary of Spodopteru frugiperdu, were investigated for their ability to grow in a serum-containing medium and in serum free media in erlenmeyer flask. An inoculum concentration of 5 × 105 cells/ml results in high growth rates in both Sf9 and Hi5 cell line. The best medium for the cell growth was sf900 SFM and Ex-cell 405 SFM in sf9 and Hi5 cell, respectively. The optimum serum-free media were selected in terms of cell density, and the selected serum free media showed higher cell growth than the medium containing FBS. References 1. Ernst-Jtirgen Schlaeger: Medium design for insect cell culture, Cytotechnology, 20, 57-70 (1996). 2. Taticek, R. A., Choi, C., Phan, S-E., Palomares, L. A., Shuler, M. L.: Comparison of Growth and Recombinant Protein Expression in Two Different Insect Cell Lines in Attached and Suspension Culture, Biotechnol. prog., 17, 676-684 (2001).
doi:10.1016/j.jbiosc.2009.08.493
AN-P34 The generation of oligodendrocytes from induced pluripotent stem cells Shinichiro Ogawa,1 Yasuhito Tokumoto,1 Jun Miyake,1,2 and Teruyuki Nagamune1
Oligodendrocytes are one type of neuronal glia cells in the central nervous system and form myelin sheaths insulating neuronal axons. Oligodendrocytes are such important cells, so the method of generating these cells has been developed for treating demyelinating disease. Recently, induced pluripotent stem (iPS) cells are generated. Although some types of cells are reported to be generated from iPS cells, the success of being differentiated into oligodendrocytes has not been reported yet. So, we tested whether iPS cells could be differentiated into oligodendrocytes by the differentiation method for ES cells and whether there were some differences in the differentiation efficiency between iPS cells and ES cells. As a result, we succeeded in generating oligodendrocytes from iPS cells. However, the differentiation efficiency of iPS cells was 2% and lower than that of ES cells. This result indicates that some factors of iPS cells inhibit differentiation and that iPS cells are not the same as ES cells. Reference 1. Yasuhito Tokumoto, Shinichiro Ogawa, Teruyuki Nagamune, Jun Miyake: “A comparison of efficiency of in vitro differentiation of oligodendrocytes from induced pluripotent stem cells versus embryonic stem cells” (in revision Experimental Cell Research, 2009).
doi:10.1016/j.jbiosc.2009.08.491
AN-P35 Study the immunomodulatory activity of Semen cuscutae and identify the active components 1
Ming-Kuem Lin, Meng-Shiou Lee, Yang,1 and Ching-Liang Chu2
Cuscuta seed, also known as Semen cuscutae or Tu-Si-Zi, is one of the commonly used Chinese medicinal materials in Asia. It is mostly used for kidney deficiency. However, the immunomodulatory activity of S. cuscutae is not known. Since dendritic cells (DCs) play a key role in regulating immune responses, we examined the effect of S. cuscutae on DCs. Mouse bone marrow-derived DCs were treated with the methanol extract of S. cuscutae (MESC), and we found that MESC induced the pro-inflammatory cytokine production by DCs, suggesting that MESC can potentially activate DCs. In addition, MESC promoted DC maturation by enhancing the expression of maturation markers. To identify the active ingredient, we further purified several compounds from MESC and tested their effect on DC activation. Unfortunately, we did not find any active candidate in first screen yet. Remarkably, our study demonstrates the immunomodulatory activity of S. cuscutae for the first time and then extends its biological function. It is interesting to reveal the molecular mechanism for this DC activation by finding the active component for further application in pharmaceutical industry. doi:10.1016/j.jbiosc.2009.08.492
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan 1 and Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka University, Osaka 560-8531, Japan 2
1
S19
1
Wen-Te Chang,
Meng-Chia
School of Chinese Medicine Resources, China Medical University, Taichung, Taiwan 1 and Immunology Research Center, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan 2
AN-P37 The production of swine cholera marker vaccine by serum free culture of insect cells infected with baculovirus Jeong Nam Park, Je Sin Woo, Chang Jin Lee, and Yeon Ho Jeong Kangwon National University, Chuncheon, Republic of Korea The development of porcine cholera marker vaccine is needed for the maintenance of porcine cholera free region. Insect cells are used as host cells for the expression of marker vaccine by baculovirus vector system. Insect cell lines are able to grow suspension culture and easy to scale up. The use of FBS has several disadvantages and is undesirable in large scale insect cell cultures media. Therefore, high density insect cell culture in serum free medium is prerequisite to produce a marker vaccine with high productivity and safety. Therefore, it is important to select the optimum serum free medium for host cell line. In this work, two insect cell lines, Hi5 originated from the eggs of Tricoplusiu ni and Sf9 derived from the ovary of Spodopteru frugiperdu, were investigated for their ability to grow in a serum-containing medium and in serum free media in erlenmeyer flask. An inoculum concentration of 5 × 105 cells/ml results in high growth rates in both Sf9 and Hi5 cell line. The best medium for the cell growth was sf900 SFM and Ex-cell 405 SFM in sf9 and Hi5 cell, respectively. The optimum serum-free media were selected in terms of cell density, and the selected serum free media showed higher cell growth than the medium containing FBS. References 1. Ernst-Jtirgen Schlaeger: Medium design for insect cell culture, Cytotechnology, 20, 57-70 (1996). 2. Taticek, R. A., Choi, C., Phan, S-E., Palomares, L. A., Shuler, M. L.: Comparison of Growth and Recombinant Protein Expression in Two Different Insect Cell Lines in Attached and Suspension Culture, Biotechnol. prog., 17, 676-684 (2001).
doi:10.1016/j.jbiosc.2009.08.493