Browse > Article
http://dx.doi.org/10.5483/BMBRep.2013.46.5.196

Morphogenetic and neuronal characterization of human neuroblastoma multicellular spheroids cultured under undifferentiated and all-trans-retinoic acid-differentiated conditions  

Jung, Gwon-Soo (Laboratory of Biochemistry and Cellular Engineering, Division of NanoBio Technology, Daegu Gyeongbuk Institute of Science and Technology)
Lee, Kyeong-Min (Laboratory of Biochemistry and Cellular Engineering, Division of NanoBio Technology, Daegu Gyeongbuk Institute of Science and Technology)
Park, Jin-Kyu (Laboratory of Biochemistry and Cellular Engineering, Division of NanoBio Technology, Daegu Gyeongbuk Institute of Science and Technology)
Choi, Seong-Kyoon (Laboratory of Biochemistry and Cellular Engineering, Division of NanoBio Technology, Daegu Gyeongbuk Institute of Science and Technology)
Jeon, Won Bae (Laboratory of Biochemistry and Cellular Engineering, Division of NanoBio Technology, Daegu Gyeongbuk Institute of Science and Technology)
Publication Information
BMB Reports / v.46, no.5, 2013 , pp. 276-281 More about this Journal
Abstract
In this study, we aimed to compare the morphogenetic and neuronal characteristics between monolayer cells and spheroids. For this purpose, we established spheroid formation by growing SH-SY5Y cells on the hydrophobic surfaces of thermally-collapsed elastin-like polypeptide. After 4 days of culture, the relative proliferation of the cells within spheroids was approximately 92% of the values for monolayer cultures. As measured by quantitative assays for mRNA and protein expressions, the production of synaptophysin and neuronspecific enolase (NSE) as well as the contents of cell adhesion molecules (CAMs) and extracellular matrix (ECM) proteins are much higher in spheroids than in monolayer cells. Under the all-trans-retinoic acid (RA)-induced differentiation condition, spheroids extended neurites and further up-regulated the expression of synaptophysin, NSE, CAMs, and ECM proteins. Our data indicate that RA-differentiated SH-SY5Y neurospheroids are functionally matured neuronal architectures.
Keywords
Cell adhesion molecules; Extracellular matrix; Multicellular neurospheroids; Neuroblastoma cells; Neuronal differentiation;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Tarnok, K., Pataki, A., Kovacs, J., Schlett, K. and Madarasz, E. (2002) Stage-dependent effects of cell-to-cell connections on in vitro induced neurogenesis. Eur. J. Cell Biol. 81, 403-412.   DOI   ScienceOn
2 Hirschhaeuser, F., Menne, H., Dittfeld, C., West, J., Mueller-Klieser, W. and Kunz-Schughart, L. A. (2010) Multicellular tumor spheroids: an underestimated tool is catching up again. J. Biotechnol. 148, 3-15.   DOI   ScienceOn
3 Ivascu, A. and Kubbies, M. (2007) Diversity of cell-mediated adhesions in breast cancer spheroids. Int. J. Oncol. 31, 1403-1413.
4 Staunton, D. E., Marlin, S. D., Stratowa, C., Dustin, M. L. and Springer, T. A. (1988) Primary structure of ICAM-I demonstrates interaction between members of the immunoglobulin and integrin supergene families. Cell 52, 925-933.   DOI   ScienceOn
5 Bao, B., Jiang, J., Yanase, T., Nishi, Y. and Morgan, J. R. Connexon-mediated cell adhesion drives microtissue self-assembly. FASEB J. 25, 255-264.
6 Bilir, A., Erguven, M., Yazihan, N., Aktas, E., Oktem, G. and Sabanci, A. (2010) Enhancement of vinorelbine-induced cytotoxicity and apoptosis by clomipramine and lithium chloride in human neuroblastoma cancer cell line SH-SY5Y. J. Neurooncol. 100, 385-395.   DOI   ScienceOn
7 Walker, K. A., Murray, T., Hilditch, T. E., Wheldon, T. E., Gregor, A. and Hann, I. M. (1988) A tumour spheroid model for antibody-targeted therapy of micrometastases. Br. J. Cancer 58, 13-16.   DOI
8 Bjerkvig, R., Lucrimi, O. D. and Rucklidge, G. J. (1989) Immunocytochemical characterization of extracellular matrix proteins expressed by cultured glioma cells. Cancer Res. 49, 5424-5428.
9 Biedler, J. L., Roffler-Tarlov, S., Schachner, M. and Freedman, L. S. (1978) Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones. Cancer Res. 38, 3751-3757.
10 Pahlman, S., Ruusala, A. I., Abrahamsson, L., Mattsson, M. E. and Esscher, T. (1984) Retinoic acid-induced differentiation of cultured human neuroblastoma cells: a comparison with phorbolester-induced differentiation. Cell Differ. 14, 135-144.   DOI   ScienceOn
11 Lopes, F. M., Schröder, R., da Frota, M. L. Jr., Zanotto-Filho, A., Müller, C. B., Pires, A. S., Meurer, R. T., Colpo, G. D., Gelain, D. P., Kapczinski, F., Moreira, J. C., Fernandes, M. C. and Klamt, F. (2010) Comparison between proliferative and neuron-like SH-SY5Y cells as an in vitro model for Parkinson disease studies. Brain Res. 1337, 85-94.   DOI   ScienceOn
12 Lombet, A., Zujovic, V., Kandouz, M., Billardon, C., Carvajal-Gonzalez, S., Gompel, A. and William, R. (2001) Resistance to induced apoptosis in the human neuroblastoma cell line SK-N-SH in relation to neuronal differentiation. Role of Bcl-2 protein family. Eur. J. Biochem. 268, 1352-1362.   DOI   ScienceOn
13 Jeon, W. B., Park, B. H., Wei, J. and Park, R. W. (2011) Stimulation of fibroblasts and neuroblasts on a biomimetic extracellular matrix consisting of tandem repeats of the elastic VGVPG domain and RGD motif. J. Biomed. Mater. Res. A 97, 2152-2157.
14 Lee, K. M., Jung, G. S., Park, J. K., Choi, S. K. and Jeon, W. B. (2013) Effects of Arg-Gly-Asp-modified elastin-like polypeptide on pseudoislet formation via up-regulation of cell adhesion molecules and extracellular matrix proteins. Acta Biomater. 9, 5600-5608.   DOI   ScienceOn
15 Lin, R. Z. and Chang, H. Y. (2008) Recent advances in three-dimensional multicellular spheroid culture for biomedical research. Biotechnol. J. 3, 1172-1184.   DOI   ScienceOn
16 Cheung, Y., Lau, W. K., Yu, M., Lai, C. S., Yeung, S., So, K. and Chang, R. C. (2009) Effects of all-trans-retinoic acid on human SH-SY5Y neuroblastoma as in vitro model in neurotoxicity research. Neurotoxicology 30, 127-135.   DOI   ScienceOn
17 Gao, X., Bian, W., Yang, J., Tang, K., Kitani, H., Atsumi, T. and Jing, N. (2001) A role of N-cadherin in neuronal differentiation of embryonic carcinoma P19 cells. Biochem. Biophys. Res. Commun. 284, 1098-1103.   DOI   ScienceOn
18 Seidenfaden, R., Krauter, A. and Hildebrandt, H. (2006) The neural cell adhesion molecule NCAM regulates neuritogenesis by multiple mechanisms of interaction. Neurochem. Int. 49, 1-11.   DOI   ScienceOn
19 Unsworth, H. C., Aasen, T., McElwaine, S. and Kelsell, D. P. (2007) Tissue-specific effects of wild-type and mutant connexin 31: a role in neurite outgrowth. Hum. Mol. Genet. 16, 165-172.   DOI
20 Rossino, P., Defilippi, P., Silengo, L. and Tarone, G. (1991) Up-regulation of the integrin $\alpha1$/$\beta1$ in human neuroblastoma cells differentiated by retinoic acid: correlation with increased neurite outgrowth response to laminin. Cell Regul. 2, 1021-1033.
21 Walker, K. A., Mairs, R., Murray, T., Hilditch, T. E., Wheldon, T. E., Gregor, A. and Hann, I. M. (1990) Tumor spheroid model for the biologically targeted radiotherapy of neuroblastoma micrometastases. Cancer Res. 50, 1000s-1002s.
22 Yuhas, J. M., Li, A. P., MartInez, A. O. and Ladman, A. J. (1997) A simplified method for production and growth of multicellular tumor spheroids. Cancer Res. 37, 3639-3643.
23 Santini, M. T., Rainaldi, G. and Indovina, P. L. (2000) Apoptosis, cell adhesion and the extracellular matrix in the three-dimensional growth of multicellular tumor spheroids. Crit. Rev. Oncol. Hematol. 36, 75-87.   DOI   ScienceOn
24 Neugebauer, K. M., Tomaselli, K. J., Lilien, J. and Reichardt, L. E. (1988) N-cadherin, NCAM, and integrins promote retinal neurite outgrowth on astrocytes in vitro. J. Cell Biol. 107, 1177-1187.   DOI