Microwell-Mediated Micro Cartilage-Like Tissue Formation of Adipose-Derived Stem Cell

Byung-Chul Kim; Jae Hyung Kim; Hyo Jung An; Wook Byun; Jae-Hong Park; Il Keun Kwon; Jin Seok Kim; Yu-Shik Hwang

Macromolecular Research, Vol.22, No.3, 287-296, March, 2014

DOI10.1007/s13233-014-2044-7 Export Citation

Microwell-Mediated Micro Cartilage-Like Tissue Formation of Adipose-Derived Stem Cell

Byung-Chul Kim, Jae Hyung Kim¹, Hyo Jung An¹, Wook Byun, Jae-Hong Park¹, Il Keun Kwon, Jin Seok Kim^{2, †}, and Yu-Shik Hwang^†

Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul, 130-701, Korea
¹Department of Pediatric Dentistry, School of Dentistry, Kyung Hee University, Seoul, 130-701, Korea
²Center for Bionics, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul, 136-791, Korea

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In cartilage tissue engineering, various technical approaches using postnatal stem cells, three-dimensional (3D) scaffolds composed of synthetic or natural polymers, and culture systems have been applied to develop 3D cartilage-like tissue. In this study, scaffold-free 3D micro-cartilage-like tissue was developed via microwell-mediated cell spheroid formation and 3D dynamic chondrogenic culture in a bioreactor. First, homogenous micro-cell spheroids were generated by the self-condensation of adipose tissue-derived stem cells (ADSCs) in microfabricated poly(ethylene glycol) (PEG) hydrogel microwells. Next, chondrogenic differentiation of the micro-ADSC spheroids was induced in the presence of transforming growth factor-beta 3 under dynamic 3D culture conditions using a high aspect ratio vessel bioreactor. Several hundred viable ADSC spheroids could be generated at a time from ADSC culture in PEG microwells. The 3D dynamic chondrogenic culture of ADSCs in the bioreactor facilitated the chondrogenic mRNA expression of proteins such as sox-9, runx2, osterix, type II collagen, and aggrecan, and the well deposition of glycosaminoglycan and type II collagen, which finally generated micro-cartilage-like tissue. Therefore, the hydrogel microwell arrays could be useful for efficiently deriving initial cell condensation-mediated chondrogenic differentiation, and for developing 3D cell-based micro-cartilage-like tissue with stem cells in a controlled manner.

Keywords:adipose tissue-derived stem cell (ADSC);condensation;chondrogenic differentiation;microwell;bioreactor;cell spheroid.

[References]

Hambly K, Bobic V, Wondrasch B, Van Assche D, Marlovits S, Am. J. Sports Med., 34, 1020 (2006)
Sterodimas A, Faria J, Nicaretta B, Pitanguy I, J. Plast.Reconstr. Aesthet. Surg., 63, 1886 (2010)
Park H, Temenoff JS, Tabata Y, Caplan AI, Mikos AG, Biomaterials., 28, 3217 (2007)
Fan J, Varshney RR, Ren L, Cai D, Wang DA, Tissue Eng. B: Rev., 15, 75 (2009)
Diekman BO, Rowland CR, Lennon DP, Caplan AI, Guilak F, Tissue Eng. A., 16, 523 (2010)
Veronesi F, Maglio M, Tschon M, Aldini N, Fini M, J. Biomed. Mater. Res. Part A., DOI: 10.1002/JBM.A.34896 (2013)
Zippel N, Schulze M, Tobiasch E, Recent Pat. Biotechnol., 4, 1 (2010)
Oliveira JT, Gardel LS, Rada T, Martins L, Gomes ME, Reis RL, J. Orthop. Res., 28, 1193 (2010)
Goldring MB, Tsuchimochi K, JiriK, J. Cell Biochem., 97, 33 (2006)
Winter A, Breit S, Parsch D, Benz K, Steck E, Hauner H, Weber RM, Ewerbeck V, Richter W, Arthritis Rheumatism., 48, 418 (2003)
Hashemibeni B, Razavi S, Esfandiary E, Karbasi S, Mardani M, Nasresfahani M, Iran. J. Basic Med. Sci., 11, 10 (2008)
Hwang YS, Chung BG, Ortmann D, Hattori N, Moeller HC, Khademhossini A, Proc. Natl. Acad. Sci., 106, 16978 (2009)
Ceuninck FD, Lesur C, Pastoureau P, Caliez A, Sabatini M, Methods Mol. Med., 100, 15 (2004)
Hwang NS, Kim MS, Sampattavanich S, Baek JH, Zhang Z, Elisseeff J, Stem Cells., 24, 284 (2006)
Poole AR, Kojima T, Yasuda T, Mwale F, Kobayashi M, Laverty S, Clin. Orthop. Relat. Res., 391, S26 (2001)
Kulyk KM, Franklin JL, Hoffman LM, Exp. Cell Res., 255, 327 (2000)
Ishaug-Riley SL, Crane-Kruger GM, Yaszemski MJ, Mikos AG, Biomaterials., 19, 1405 (1998)
Glowacki J, Mizuno S, Greenberger JS, Cell Transplant., 7, 319 (1998)
Holy CE, Shoichet MS, Davies JE, J. Biomed. Mater.Res., 51, 376 (2000)
Mueller SM, Mizuno S, Gerstenfeld LC, Glowacki J, J. Bone Miner. Res., 14, 2118 (1999)
Allison P. Van W, Gates ID, Kallos SM, Cells Tissues Organs., 196, 34 (2012)
Hwang YS, Cho J, Chan KLA, Heng J, Boccacini AR, Kazarian SG, Williams D, Polak JM, Mantalaris A, Biomaterials., 30, 499 (2009)

[Referenced By]

[1] Hambly K, Bobic V, Wondrasch B, Van Assche D, Marlovits S, Am. J. Sports Med., 34, 1020 (2006)

[2] Sterodimas A, Faria J, Nicaretta B, Pitanguy I, J. Plast.Reconstr. Aesthet. Surg., 63, 1886 (2010)

[3] Park H, Temenoff JS, Tabata Y, Caplan AI, Mikos AG, Biomaterials., 28, 3217 (2007)

[4] Fan J, Varshney RR, Ren L, Cai D, Wang DA, Tissue Eng. B: Rev., 15, 75 (2009)

[5] Diekman BO, Rowland CR, Lennon DP, Caplan AI, Guilak F, Tissue Eng. A., 16, 523 (2010)

[6] Veronesi F, Maglio M, Tschon M, Aldini N, Fini M, J. Biomed. Mater. Res. Part A., DOI: 10.1002/JBM.A.34896 (2013)

[7] Zippel N, Schulze M, Tobiasch E, Recent Pat. Biotechnol., 4, 1 (2010)

[8] Oliveira JT, Gardel LS, Rada T, Martins L, Gomes ME, Reis RL, J. Orthop. Res., 28, 1193 (2010)

[9] Goldring MB, Tsuchimochi K, JiriK, J. Cell Biochem., 97, 33 (2006)

[10] Winter A, Breit S, Parsch D, Benz K, Steck E, Hauner H, Weber RM, Ewerbeck V, Richter W, Arthritis Rheumatism., 48, 418 (2003)

[11] Hashemibeni B, Razavi S, Esfandiary E, Karbasi S, Mardani M, Nasresfahani M, Iran. J. Basic Med. Sci., 11, 10 (2008)

[12] Hwang YS, Chung BG, Ortmann D, Hattori N, Moeller HC, Khademhossini A, Proc. Natl. Acad. Sci., 106, 16978 (2009)

[13] Ceuninck FD, Lesur C, Pastoureau P, Caliez A, Sabatini M, Methods Mol. Med., 100, 15 (2004)

[14] Hwang NS, Kim MS, Sampattavanich S, Baek JH, Zhang Z, Elisseeff J, Stem Cells., 24, 284 (2006)

[15] Poole AR, Kojima T, Yasuda T, Mwale F, Kobayashi M, Laverty S, Clin. Orthop. Relat. Res., 391, S26 (2001)

[16] Kulyk KM, Franklin JL, Hoffman LM, Exp. Cell Res., 255, 327 (2000)

[17] Ishaug-Riley SL, Crane-Kruger GM, Yaszemski MJ, Mikos AG, Biomaterials., 19, 1405 (1998)

[18] Glowacki J, Mizuno S, Greenberger JS, Cell Transplant., 7, 319 (1998)

[19] Holy CE, Shoichet MS, Davies JE, J. Biomed. Mater.Res., 51, 376 (2000)

[20] Mueller SM, Mizuno S, Gerstenfeld LC, Glowacki J, J. Bone Miner. Res., 14, 2118 (1999)

[21] Allison P. Van W, Gates ID, Kallos SM, Cells Tissues Organs., 196, 34 (2012)

[22] Hwang YS, Cho J, Chan KLA, Heng J, Boccacini AR, Kazarian SG, Williams D, Polak JM, Mantalaris A, Biomaterials., 30, 499 (2009)