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Development of three-dimensional perfusion system using microbioreactor for stem cell culture
S182
Abstracts / Journal of Biotechnology 136S (2008) S176–S182
References Ballen, K.K., 2005. New trends in umbilical cord blood transplantation. Blood 105, 3786–3792. Wagner, W., Roderburg, C., Wein, F., Diehmann, A., Frankhauser, M., Schubert, R., Eckstein, V., Ho, A.D., 2007. Molecular and secretory profiles of human mesenchymal stromal cells and their abilities to maintain primitive hematopoietic progenitors. Stem Cells 25, 2638–2647. Zhang, Y., Li, C., Jiang, X., Zhang, S., Wu, Y., Liu, B., Tang, P., Mao, N., 2004. Human placenta-derived mesenchymal progenitor cells support culture expansion of long-term culture-initiating cells from cord blood CD34+ cells. Exp. Hematol. 32, 657–664.
doi:10.1016/j.jbiotec.2008.07.401 III1-YP-014 In vitro cardiomyogenic potential of human periosteumderived progenitor cells Jong-Il Kim 1 , Ji-Hae Song 1 , Bu-Young Park 1 , Yong-Soo Choi 1 , Hyung-Jin Jeon 1 , Young-Joo Jeon 1 , Mi-Eun Gong 1 , Sang-Min Lim 2 , Chang-Woo Lee 3 , Dong-Il Kim 1,∗ 1
Department of Biological Engineering, Inha University, Incheon, Republic of Korea 2 Central Research Institute, Boryung Pharmaceutical Co. Ltd., Ansan, Republic of Korea 3 Good Shepherd Hospital, Yeoksam-Dong, Seoul, Republic of Korea E-mail address: [email protected] (D.-I. Kim). Regeneration of myocadiac tissue is limited in adult life and is not sufficient to compensate for cell loss with myocardial infarction. Therefore, the identification of a putative source of cardiomyocyte progenitor cells is of great interest to provide a usable model in vitro (Planat-Benard et al., 2004). Among various mesodermal tissues, periosteum is relatively easy to access and contains multipotent mesenchymal stem cells (Choi et al., 2007). In this study, the potential of human periosteum-derived progenitor cells (PDPCs) to differentiate into a cardiomyocyte phenotype was investigated. PDPCs were identified by the expression of many molecules including CD105 (SH2) and CD73 (SH3/4), and were negative for the hematopoietic markers CD34, CD45 and CD14. PDPCs were induced with 5-azacytidine for 24 h (Makino et al., 1999). Then incubated with TGF1, IGF1 in Maintenance media. After 5 weeks later, induced PDPCs were analyzed by morphological observation and histological staining. The expression of cardiomyocyte-specific mRNA such as GATA-4, Nkx2.5 was verified by RT-PCR analysis. These results indicate that the PDPCs can be used as an alternative cell source for autologous myocardial repair. References Choi, Y.S., Lim, S.M., Shin, H.C., Lee, C.W., Kim, D.I., 2007. Chondrogenesis of human periosteum-derived progenitor cells in atelocollagen. Biotechnol. Lett. 29, 323–329.
Makino, S., Fukuda, K., Miyoshi, S., 1999. Cardiomyocytes can be generated from marrow stromal cells in vitro. J. Clin. Invest. 103, 697–705. Planat-Benard, V., Menard, C., Andre, M., Puceat, M., Perez, A., Garcia-Verdugo, J.M., Penicaud, L., Casteilla, L., 2004. Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells. Circ. Res. 94, 223–229.
doi:10.1016/j.jbiotec.2008.07.402 III1-YP-015 Development of three-dimensional perfusion system using microbioreactor for stem cell culture Ji-Hae Song 1 , Yong-Soo Choi 1 , Jong-Il Kim 1 , Bu-Young Park 1 , Hyung-Jin Jeon 1 , Young-Joo Jeon 1 , Mi-Eun Kong 1 , Kyu-Ho Chang 2 , Dong-Il Kim 1,∗ 1
Department of Biological Engineering, Inha University, Incheon, Republic of Korea 2 Biotron Inc., Bucheon, Republic of Korea E-mail address: [email protected] (D.-I. Kim). Mesenchymal stem cells (MSCs) cultured in three-dimensional (3D) scaffold can be used for the repair of articular cartilage defects. Perfusion system in bioreactor has beneficial effects on cell capacity and tissue growth since it improves the nutrient supply and the exchange of wastes. A microbioreactor for the perfusion cultures of stem cells was developed so that MSCs can be induced to differentiate into chondrocytes. MSCs isolated from umbilical cord blood were seeded and cultured within a bovine collagen sponge composed of extracellular matrix secreted from foreskin-derived fibroblasts. Chondrogenic differentiation of MSCs was induced in the microbioreactor for 3 weeks. The location of MSCs in the scaffold could be observed by scanning electron microscope. Chondrogenic potential of MSCs was verified by histological staining and RT-PCR. The cells within the 3D scaffold in perfusion system were localized to the central part rather than the construct periphery in contrast to the static culture. Moreover, MSCs could be successfully differentiated into chondrocytes in the microbioreactor than those in 2D culture. These results show that the perfusion culture in microbioreactor can offer articular cartilage-like tissues. References Xu, S., Li, D., Xie, Y., Lu, J., Dai, K., 2007. The growth of stem cells within beta-TCP scaffolds in a fluid-dynamic environment. Mater. Sci. Eng. C 28, 164–170. Zhao, F., Chella, R., Ma, T., 2007. Effects of shear stress on 3-D human mesenchymal stem cell construct development in a perfusion bioreactor system: experiments and hydrodynamic modeling. Biotechnol. Bioeng. 96, 584–595.
doi:10.1016/j.jbiotec.2008.07.403
Abstracts / Journal of Biotechnology 136S (2008) S176–S182
References Ballen, K.K., 2005. New trends in umbilical cord blood transplantation. Blood 105, 3786–3792. Wagner, W., Roderburg, C., Wein, F., Diehmann, A., Frankhauser, M., Schubert, R., Eckstein, V., Ho, A.D., 2007. Molecular and secretory profiles of human mesenchymal stromal cells and their abilities to maintain primitive hematopoietic progenitors. Stem Cells 25, 2638–2647. Zhang, Y., Li, C., Jiang, X., Zhang, S., Wu, Y., Liu, B., Tang, P., Mao, N., 2004. Human placenta-derived mesenchymal progenitor cells support culture expansion of long-term culture-initiating cells from cord blood CD34+ cells. Exp. Hematol. 32, 657–664.
doi:10.1016/j.jbiotec.2008.07.401 III1-YP-014 In vitro cardiomyogenic potential of human periosteumderived progenitor cells Jong-Il Kim 1 , Ji-Hae Song 1 , Bu-Young Park 1 , Yong-Soo Choi 1 , Hyung-Jin Jeon 1 , Young-Joo Jeon 1 , Mi-Eun Gong 1 , Sang-Min Lim 2 , Chang-Woo Lee 3 , Dong-Il Kim 1,∗ 1
Department of Biological Engineering, Inha University, Incheon, Republic of Korea 2 Central Research Institute, Boryung Pharmaceutical Co. Ltd., Ansan, Republic of Korea 3 Good Shepherd Hospital, Yeoksam-Dong, Seoul, Republic of Korea E-mail address: [email protected] (D.-I. Kim). Regeneration of myocadiac tissue is limited in adult life and is not sufficient to compensate for cell loss with myocardial infarction. Therefore, the identification of a putative source of cardiomyocyte progenitor cells is of great interest to provide a usable model in vitro (Planat-Benard et al., 2004). Among various mesodermal tissues, periosteum is relatively easy to access and contains multipotent mesenchymal stem cells (Choi et al., 2007). In this study, the potential of human periosteum-derived progenitor cells (PDPCs) to differentiate into a cardiomyocyte phenotype was investigated. PDPCs were identified by the expression of many molecules including CD105 (SH2) and CD73 (SH3/4), and were negative for the hematopoietic markers CD34, CD45 and CD14. PDPCs were induced with 5-azacytidine for 24 h (Makino et al., 1999). Then incubated with TGF1, IGF1 in Maintenance media. After 5 weeks later, induced PDPCs were analyzed by morphological observation and histological staining. The expression of cardiomyocyte-specific mRNA such as GATA-4, Nkx2.5 was verified by RT-PCR analysis. These results indicate that the PDPCs can be used as an alternative cell source for autologous myocardial repair. References Choi, Y.S., Lim, S.M., Shin, H.C., Lee, C.W., Kim, D.I., 2007. Chondrogenesis of human periosteum-derived progenitor cells in atelocollagen. Biotechnol. Lett. 29, 323–329.
Makino, S., Fukuda, K., Miyoshi, S., 1999. Cardiomyocytes can be generated from marrow stromal cells in vitro. J. Clin. Invest. 103, 697–705. Planat-Benard, V., Menard, C., Andre, M., Puceat, M., Perez, A., Garcia-Verdugo, J.M., Penicaud, L., Casteilla, L., 2004. Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells. Circ. Res. 94, 223–229.
doi:10.1016/j.jbiotec.2008.07.402 III1-YP-015 Development of three-dimensional perfusion system using microbioreactor for stem cell culture Ji-Hae Song 1 , Yong-Soo Choi 1 , Jong-Il Kim 1 , Bu-Young Park 1 , Hyung-Jin Jeon 1 , Young-Joo Jeon 1 , Mi-Eun Kong 1 , Kyu-Ho Chang 2 , Dong-Il Kim 1,∗ 1
Department of Biological Engineering, Inha University, Incheon, Republic of Korea 2 Biotron Inc., Bucheon, Republic of Korea E-mail address: [email protected] (D.-I. Kim). Mesenchymal stem cells (MSCs) cultured in three-dimensional (3D) scaffold can be used for the repair of articular cartilage defects. Perfusion system in bioreactor has beneficial effects on cell capacity and tissue growth since it improves the nutrient supply and the exchange of wastes. A microbioreactor for the perfusion cultures of stem cells was developed so that MSCs can be induced to differentiate into chondrocytes. MSCs isolated from umbilical cord blood were seeded and cultured within a bovine collagen sponge composed of extracellular matrix secreted from foreskin-derived fibroblasts. Chondrogenic differentiation of MSCs was induced in the microbioreactor for 3 weeks. The location of MSCs in the scaffold could be observed by scanning electron microscope. Chondrogenic potential of MSCs was verified by histological staining and RT-PCR. The cells within the 3D scaffold in perfusion system were localized to the central part rather than the construct periphery in contrast to the static culture. Moreover, MSCs could be successfully differentiated into chondrocytes in the microbioreactor than those in 2D culture. These results show that the perfusion culture in microbioreactor can offer articular cartilage-like tissues. References Xu, S., Li, D., Xie, Y., Lu, J., Dai, K., 2007. The growth of stem cells within beta-TCP scaffolds in a fluid-dynamic environment. Mater. Sci. Eng. C 28, 164–170. Zhao, F., Chella, R., Ma, T., 2007. Effects of shear stress on 3-D human mesenchymal stem cell construct development in a perfusion bioreactor system: experiments and hydrodynamic modeling. Biotechnol. Bioeng. 96, 584–595.
doi:10.1016/j.jbiotec.2008.07.403