Fabrication of micrometer-scale porous gelatin scaffolds for 3D cell culture

Kyuyoung Shim; So Hyun Kim; Dongwook Lee; Bumsang Kim; Tae Hee Kim; Youngmee Jung; Nakwon Choi; Jong Hwan Sung

Journal of Industrial and Engineering Chemistry, Vol.50, 183-189, June, 2017

DOI10.1016/j.jiec.2017.02.012 Export Citation

Fabrication of micrometer-scale porous gelatin scaffolds for 3D cell culture

Kyuyoung Shim¹, So Hyun Kim^{2, 3}, Dongwook Lee¹, Bumsang Kim¹, Tae Hee Kim^{4, 5}, Youngmee Jung^{5, 6}, Nakwon Choi^{2, 6, †}, and Jong Hwan Sung^{1, †}

¹Department of Chemical Engineering, Hongik University, Seoul, Republic of Korea
²Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
³Specialty Medicine Team, SK Biopharmaceuticals Co., Ltd., Republic of Korea
⁴NBIT, KU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul, Republic of Korea
⁵Center for Biomaterials, Biomedical Research Institute, Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
⁶Department of Biomedical Engineering, Korea University of Science and Technology (UST), Daejeon, Republic of Korea

E-mail:,

In hydrogel scaffolds for 3D cell culture, porous structure plays a central role in maintaining cell survival by delivery of biochemical factors through the scaffolds. Thus, adequate pore structures should be a significant design factor for preparing cell culture scaffolds. Here, we present a non cytotoxic approach to control porosity of a gelatin scaffold based on porogen leaching. Specifically, we fabricated photocrosslinkable hydrogel (gelatin methacrylate) containing porogens in a microfluidic channel, followed by dissolving out the porogens. Our approach allowed generation of micrometer scale pores, increasing cell viability. This study could serve as a guide for tuning porosity of cell-seeded 3D scaffolds.

Keywords:Gelatin-methacrylate (Gel-MA);Micrometer-scale porous hydrogel scaffold;Photocrosslinking;Porogen;3D cell culture

[References]

Costello CM, Jia HP, Shaffiey S, Yu JJ, Jain NK, Hackam D, March JC, Biotechnol. Bioeng., 111(6), 1222 (2014)
Kim HJ, Huh D, Hamilton G, Ingber DE, Lab Chip, 12, 2165 (2012)
Pusch J, Votteler M, Gohler S, Engl J, Hampel M, Walles H, Schenke- Layland K, Biomaterials, 32, 7469 (2011)
Lee SA, No DY, Kang E, Ju J, Kim DS, Lee SH, Lab Chip, 13, 3529 (2013)
Bellas E, Seiberg M, Garlick J, Kaplan DL, Macromol. Biosci., 12, 1627 (2012)
Gangatirkar P, Paquet-Fifield S, Li A, Rossi R, Kaur P, Nat. Protoc., 2, 178 (2007)
Miller JS, Stevens KR, Yang MT, Baker BM, Nguyen DHT, Cohen DM, Toro E, Chen AA, Galie PA, Yu X, Chaturvedi R, Bhatia SN, Chen CS, Nat. Mater., 11(9), 768 (2012)
Peppas NA, Bures P, Leobandung W, Ichikawa H, Eur. J. Pharm. Biopharm., 50, 27 (2000)
Rosales AM, Anseth KS, Nat. Rev. Mater., 1 (2016)
Choi NW, Cabodi M, Held B, Gleghorn JP, Bonassar LJ, Stroock AD, Nat. Mater., 6(11), 908 (2007)
Cabodi M, Choi NW, Gleghorn JP, Lee CSD, Bonassar LJ, Stroock AD, J. Am. Chem. Soc., 127(40), 13788 (2005)
Sung JH, Yu JJ, Luo D, Shuler MJ, March JC, Lab Chip, 11, 389 (2011)
Kuo CK, Ma PX, Biomaterials, 22, 511 (2001)
Chen L, Subirade M, Biomaterials, 27, 4646 (2006)
Freiberg S, Zhu XX, Int. J. Pharm., 282, 1 (2004)
Cross VL, Zheng Y, Choi NW, Verbridge SS, Sutermaster BA, Bonassar LJ, Fischbach C, Stroock AD, Biomaterials, 31, 8596 (2010)
Kim JA, Kim HN, Im SK, Chung S, Kang JY, Choi N, Biomicrofluidics, 9 (2015)
Morgan JP, Delnero PF, Zheng Y, Verbridge SS, Chen JM, Craven M, Choi NW, Diaz-Santana A, Kermani P, Hempstead B, Lopez JA, Corso TN, Fischbach C, Stroock AD, Nat. Protoc., 8, 1820 (2013)
Zheng Y, Chen JM, Craven M, Choi NW, Totorica S, Diaz-Santana A, Kermani P, Hempstead B, Fischbach-Teschl C, Lopez JA, Stroock AD, Proc. Natl. Acad. Sci. U. S. A., 109, 9342 (2012)
Forgacs G, Newman SA, Hinner B, Maier CW, Sackmann E, Biophys. J., 84, 1272 (2003)
Kang HW, Tabata Y, Ikada Y, Biomaterials, 20, 1339 (1999)
Lin CC, Anseth KS, Pharm. Res., 26, 631 (2009)
Weber LW, He J, Bradley B, Haskins K, Anseth KS, Acta Biomater., 2, 1 (2006)
Peppas NA, Hilt JZ, Khademhosseini A, Langer R, Adv. Mater., 18(11), 1345 (2006)
Astete CE, Sabliov CM, J. Biomater. Sci.-Polym. Ed., 17, 247 (2006)
Park TG, Biomaterials, 16, 1123 (1995)
Bruzauskaite I, Bironaite D, Bagdonas E, Bernotiene E, Cytotechnology, 68, 355 (2016)
Lips PAM, Velthoen IW, Dijkstra PJ, Wessling M, Feijen J, Polymer, 46(22), 9396 (2005)
Yang SF, Leong KF, Du ZF, Chua CK, Tissue Eng., 7, 679 (2001)
Stokols S, Tuszynski MH, Biomaterials, 25, 5839 (2004)
Simms HM, Brotherton CM, Good BT, Davis RH, Anseth KS, Bowman CN, Lab Chip, 5, 151 (2005)
Han LH, Lai JH, Yu S, Yang F, Biomaterials, 34, 4251 (2013)
Hwang CM, Sant S, Masaeli M, Kachouie NN, Zamanian B, Lee SH, Khademhosseini A, Biofabrication, 2 (2010)
Kim J, Yaszemski MJ, Lu LC, Tissue Eng., 15, 583 (2009)
Scott EA, Nichols MD, Kuntz-Willits R, Elbert DL, Acta Biomater., 6, 29 (2010)
Gonzalez-Meijome JM, Lopez-Alemany A, Almeida JB, Parafita MA, Refojo MF, J. Biomed. Mater. Res., 76, 419 (2006)
Fournier RL, Basic Transport Phenomena in Biomedical Engineering, 2nd ed., Taylor & Francis, New York, 2007.
Choi NW, Kim J, Chapin SC, Duong T, Donohue E, Pandey P, Broom W, Hill WA, Doyle PS, Anal. Chem., 84, 9370 (2012)
Lee AG, Arena CP, Beebe DJ, Palecek SP, Biomacromolecules, 11(12), 3316 (2010)
Ma PX, Adv. Drug Deliv. Rev., 60, 184 (2008)
Williams CG, Malik AN, Kim TK, Manson PN, Elisseeff JH, Biomaterials, 26, 1211 (2005)
Jung J, Oh J, Dig. J. Nanomater. Biostruct., 9, 503 (2014)
Chen MB, Srigunapalan S, Wheeler AR, Simmons CA, Lab Chip, 13, 2013 (2591)
Gong Y, Ma Z, Zhou Q, Li J, Gao C, Shen J, J. Biomater. Sci.-Polym. Ed., 19, 207 (2008)
Cisewski SE, Zhang L, Kuo J, Wright GJ, Wu Y, Kern MJ, Yao H, Osteoarthritis Cartilage, 23, 1790 (2015)

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[23] Weber LW, He J, Bradley B, Haskins K, Anseth KS, Acta Biomater., 2, 1 (2006)

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