Size Regulation of Chondrocyte Spheroids Using a PDMS-Based Cell Culture Chip

Takahisa Anada; Osamu Suzuki

doi:10.20965/jrm.2013.p0644

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JRM Vol.25 No.4 pp. 644-649

doi: 10.20965/jrm.2013.p0644

(2013)

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Size Regulation of Chondrocyte Spheroids Using a PDMS-Based Cell Culture Chip

Takahisa Anada and Osamu Suzuki

Division of Craniofacial Function Engineering, Graduate School of Dentistry, Tohoku University, 4-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan

Received:

January 31, 2013

Accepted:

June 4, 2013

Published:

August 20, 2013

Keywords:

chondrocyte, spheroid, culture chip

Abstract

Cartilage self-repair is limited due to a lack of blood supply and the low mitosis rate of chondrocytes. A tissue engineering approach using cells and biomaterials has the potential to treat cartilage injury. Threedimensional cellular aggregates are an excellent model for mimicking condensation and chondrogenic differentiation in vitro. We developed a technique for constructing spheroids utilizing a polydimethylsiloxane (PDMS)-based culture chip. The objective of this study is to determine how the initial cell density on a culture chip affects the chondrogenic ATDC5 cell differentiation. We demonstrate how culture chips having arrays of multicavities are able to generate high numbers of uniform spheroids rapidly and simultaneously with narrow size distribution. Spheroids are collected easily and noninvasively. Higher cell seeding density on the culture chip enhances chondrogenic cell differentiation. These results suggest the usefulness of this chip in engineering 3D cellular constructs with high functionality for tissue engineering.

Cite this article as:

T. Anada and O. Suzuki, “Size Regulation of Chondrocyte Spheroids Using a PDMS-Based Cell Culture Chip,” J. Robot. Mechatron., Vol.25 No.4, pp. 644-649, 2013.

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References

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[28] K. Gelse, M. Brem, P. Klinger, A. Hess, B. Swoboda, F. Hennig et al., “Paracrine effect of transplanted rib chondrocyte spheroids supports formation of secondary cartilage repair tissue,” J. of orthopaedic research: official publication of the Orthopaedic Research Society, Vol.27, No.9, pp. 1216-1225, 2009.
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This article is published under a Creative Commons Attribution-NoDerivatives 4.0 Internationa License.

[1] [1] V. Mironov, R. P. Visconti, V. Kasyanov, G. Forgacs, C. J. Drake, and R. R. Markwald, “Organ printing: tissue spheroids as building blocks,” Biomaterials, Vol.30, No.12, pp. 2164-2174, 2009.

[2] [2] L. G. Griffith and M. A. Swartz, “Capturing complex 3D tissue physiology in vitro,” Nature reviews Molecular cell biology, Vol.7, No.3, pp. 211-224, 2006.

[3] [3] J. M. Kelm, N. E. Timmins, C. J. Brown, M. Fussenegger, and L. K. Nielsen, “Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types,” Biotechnol. Bioeng., Vol.83, No.2, pp. 173-180, 2003.

[4] [4] L. A. Kunz-Schughart, M. Kreutz, and R. Knuechel, “Multicellular spheroids: a three-dimensional in vitro culture system to study tumour biology,” Int. J. Exp. Pathol., Vol.79, No.1, pp. 1-23, 1998.

[5] [5] K. Yamada, M. Kamihira, R. Hamamoto, and S. Iijima, “Efficient induction of hepatocyte spheroids in a suspension culture using a water-soluble synthetic polymer as an artificial matrix,” J. Biochem., Vol.123, No.6, pp. 1017-1023, 1998.

[6] [6] E. S. Ng, R. P. Davis, L. Azzola, E. G. Stanley, and A. G. Elefanty, “Forced aggregation of defined numbers of human embryonic stem cells into embryoid bodies fosters robust, reproducible hematopoietic differentiation,” Blood, Vol.106, No.5, pp. 1601-1603, 2005.

[7] [7] A. Asthana and W. S. Kisaalita, “Microtissue size and hypoxia in HTS with 3D cultures,” Drug discovery today, 2012.

[8] [8] Y. S. Hwang, B. G. Chung, D. Ortmann, N. Hattori, H. C. Moeller, and A. Khademhosseini, “Microwell-mediated control of embryoid body size regulates embryonic stem cell fate via differential expression of WNT5a and WNT11,” Proc. of the National Academy of Sciences of the United States of America, Vol.106, No.40, pp. 16978-16983, 2009.

[9] [9] K. Kikuchi, K. Sumaru, J. Edahiro, Y. Ooshima, S. Sugiura, T. Takagi et al., “Stepwise assembly of micropatterned co-cultures using photoresponsive culture surfaces and its application to hepatic tissue arrays,” Biotechnol. Bioeng., Vol.103, No.3, pp. 552-561, 2009.

[10] [10] Y. Sakai, Y. Yoshiura, and K. Nakazawa, “Embryoid body culture of mouse embryonic stem cells using microwell and micropatterned chips,” J. Biosci. Bioeng., Vol.111, No.1, pp. 85-91, 2011.

[11] [11] Y. S. Torisawa, B. H. Chueh, D. Huh, P. Ramamurthy, T. M. Roth, K. F. Barald et al., “Efficient formation of uniform-sized embryoid bodies using a compartmentalized microchannel device,” Lab Chip, Vol.7, No.6, pp. 770-776, 2007.

[12] [12] W. Wang, K. Itaka, S. Ohba, N. Nishiyama, U. I. Chung, Y. Yamasaki et al., “3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells,” Biomaterials, Vol.30, No.14, pp. 2705-2715, 2009.

[13] [13] H. M. Kronenberg, “Developmental regulation of the growth plate,” Nature, Vol.423, No.6937, pp. 332-336, 2003.

[14] [14] T. Atsumi, Y.Miwa, K. Kimata, and Y. Ikawa, “A chondrogenic cell line derived from a differentiating culture of AT805 teratocarcinoma cells,” Cell differentiation and development: the official journal of the Int. Society of Developmental Biologists, Vol.30, No.2, pp. 109-116, 1990.

[15] [15] C. Shukunami, K. Ishizeki, T. Atsumi, Y. Ohta, F. Suzuki, and Y. Hiraki, “Cellular hypertrophy and calcification of embryonal carcinoma-derived chondrogenic cell line ATDC5 in vitro,” J. of bone and mineral research: the official journal of the American Society for Bone and Mineral Research, Vol.12, No.8, pp. 1174-1188, 1997.

[16] [16] T. Anada, J. Fukuda, Y. Sai, and O. Suzuki, “An oxygen-permeable spheroid culture system for the prevention of central hypoxia and necrosis of spheroids,” Biomaterials, Vol.33, No.33, pp. 8430-8441, 2012.

[17] [17] T. Anada, T. Masuda, Y. Honda, J. Fukuda, F. Arai, T. Fukuda et al., “Three-dimensional cell culture device utilizing thin membrane deformation by decompression,” Sensors and Actuators B: Chemical, Vol.147, No.1, pp. 376-379, 2010.

[18] [18] S. Sugiura, J. Edahiro, K. Sumaru, and T. Kanamori, “Surface modification of polydimethylsiloxane with photo-grafted poly(ethylene glycol) for micropatterned protein adsorption and cell adhesion,” Colloids and surfaces B, Biointerfaces, Vol.63, No.2, pp. 301-305, 2008.

[19] [19] J. C. McDonald and G. M. Whitesides, “Poly(dimethylsiloxane) as a material for fabricating microfluidic devices,” Acc. Chem. Res., Vol.35, pp. 491-499, 2002.

[20] [20] J. M. Karp, J. Yeh, G. Eng, J. Fukuda, J. Blumling, K. Y. Suh et al., “Controlling size, shape and homogeneity of embryoid bodies using poly(ethylene glycol) microwells,” Lab Chip, Vol.7, No.6, pp. 786-794, 2007.

[21] [21] M. Takagi, Y. Umetsu, M. Fujiwara, and S. Wakitani, “High inoculation cell density could accelerate the differentiation of human bone marrow mesenchymal stem cells to chondrocyte cells,” J. Biosci. Bioeng., Vol.103, No.1, pp. 98-100, 2007.

[22] [22] T. Y. Hui, K. M. Cheung, W. L. Cheung, D. Chan, and B. P. Chan, “In vitro chondrogenic differentiation of human mesenchymal stem cells in collagen microspheres: influence of cell seeding density and collagen concentration,” Biomaterials, Vol.29, No.22, pp. 3201-3212, 2008.

[23] [23] C. M. Revell, C. E. Reynolds, and K. A. Athanasiou, “Effects of initial cell seeding in self assembly of articular cartilage. Annals of biomedical engineering,” Vol.36, No.9, pp. 1441-1448, 2008.

[24] [24] A. M. Delise and R. S. Tuan, “Analysis of N-cadherin function in limb mesenchymal chondrogenesis in vitro,” Developmental dynamics: an official publication of the American Association of Anatomists, Vol.225, No.2, pp. 195-204, 2002.

[25] [25] A. B. Adesida, A. Mulet-Sierra, and N. M. Jomha, “Hypoxia mediated isolation and expansion enhances the chondrogenic capacity of bone marrow mesenchymal stromal cells,” Stem cell research & therapy, Vol.3, No.2, p. 9, 2012.

[26] [26] M. Hirao, N. Tamai, N. Tsumaki, H. Yoshikawa, and A. Myoui, “Oxygen tension regulates chondrocyte differentiation and function during endochondral ossification,” The J. of biological chemistry, Vol.281, No.41, pp. 31079-31092, 2006.

[27] [27] S. Strobel, M. Loparic, D. Wendt, A. D. Schenk, C. Candrian, R. L. Lindberg et al., “Anabolic and catabolic responses of human articular chondrocytes to varying oxygen percentages,” Arthritis research & therapy, Vol.12, No.2, p. R34, 2010.

[28] [28] K. Gelse, M. Brem, P. Klinger, A. Hess, B. Swoboda, F. Hennig et al., “Paracrine effect of transplanted rib chondrocyte spheroids supports formation of secondary cartilage repair tissue,” J. of orthopaedic research: official publication of the Orthopaedic Research Society, Vol.27, No.9, pp. 1216-1225, 2009.

[29] [29] J. I. Lee, M. Sato, H. W. Kim, and J. Mochida, “Transplantation of scaffold-free spheroids composed of synovium-derived cells and chondrocytes for the treatment of cartilage defects of the knee,” European cells & materials, Vol.22, pp. 275-290, discussion 90, 2011.

[30] [30] T. Schubert, S. Anders, E. Neumann, J. Scholmerich, F. Hofstadter, J. Grifka et al., “Long-term effects of chondrospheres on cartilage lesions in an autologous chondrocyte implantation model as investigated in the SCID mouse model,” Int. J. Mol. Med., Vol.23, No.4, pp. 455-460, 2009.

[31] [31] H. H. Yoon, S. H. Bhang, J. Y. Shin, J. Shin, and B. S. Kim, “Enhanced cartilage formation via three-dimensional cell engineering of human adipose-derived stem cells,” Tissue engineering Part A, Vol.18, No.19-20, pp. 1949-1956, 2012.