[en] Human pluripotent stem cells hold great promise for applications in drug discovery and regenerative medicine. Microfluidic technology is a promising approach for creating artificial microenvironments; however, although a proper 3D microenvironment is required to achieve robust control of cellular phenotypes, most current microfluidic devices provide only 2D cell culture and do not allow tuning of physical and chemical environmental cues simultaneously. Here, the authors report a 3D cellular microenvironment plate (3D-CEP), which consists of a microfluidic device filled with thermoresponsive poly(N-isopropylacrylamide)-beta-poly(ethylene glycol) hydrogel (HG), which enables systematic tuning of both chemical and physical environmental cues as well as in situ cell monitoring. The authors show that H9 human embryonic stem cells (hESCs) and 253G1 human induced pluripotent stem cells in the HG/3D-CEP system maintain their pluripotent marker expression under HG/3D-CEP self-renewing conditions. Additionally, global gene expression analyses are used to elucidate small variations among different test environments. Interestingly, the authors find that treatment of H9 hESCs under HG/3D-CEP self-renewing conditions results in initiation of entry into the neural differentiation process by induction of PAX3 and OTX1 expression. The authors believe that this HG/3D-CEP system will serve as a versatile platform for developing targeted functional cell lines and facilitate advances in drug screening and regenerative medicine.
Disciplines :
Biotechnology
Author, co-author :
Kamei, Ken-Ichiro
Koyama, Yoshie
Tokunaga, Yumie ; Université de Liège - ULiège > Medical Genomics-Unit of Animal Genomics
Mashimo, Yasumasa
Yoshioka, Momoko
Fockenberg, Christopher
Mosbergen, Rowland
Korn, Othmar
Wells, Christine
Chen, Yong
Language :
English
Title :
Characterization of Phenotypic and Transcriptional Differences in Human Pluripotent Stem Cells under 2D and 3D Culture Conditions.
J. A. Thomson, J. Itskovitz-Eldor, S. S. Shapiro, M. A. Waknitz, J. J. Swiergiel, V. S. Marshall, J. M. Jones, Science 1998, 282, 1145.
K. Takahashi, K. Tanabe, M. Ohnuki, M. Narita, T. Ichisaka, K. Tomoda, S. Yamanaka, Cell 2007, 131, 861;
J. Yu, M. A. Vodyanik, K. Smuga-Otto, J. Antosiewicz-Bourget, J. L. Frane, S. Tian, J. Nie, G. A. Jonsdottir, V. Ruotti, R. Stewart, I. I. Slukvin, J. A. Thomson, Science 2007, 318, 1917;
W. E. Lowry, L. Richter, R. Yachechko, A. D. Pyle, J. Tchieu, R. Sridharan, A. T. Clark, K. Plath, Proc. Natl. Acad. Sci. USA 2008, 105, 2883.
D. E. Discher, D. J. Mooney, P. W. Zandstra, Science 2009, 324, 1673;
D. Nampe, H. Tsutsui, J. Lab. Autom. 2013, 18, 482;
S. W. Lane, D. A. Williams, F. M. Watt, Nat. Biotechnol. 2014, 32, 795.
K. Kamei, J. Lab. Autom. 2013, 18, 469;
O. Gagliano, N. Elvassore, C. Luni, Biochem. Biophys. Res. Commun. 2016, 473, 683;
T. Qian, E. V. Shusta, S. P. Palecek, Curr. Opin. Genet. Dev. 2015, 34, 54;
P. Ertl, D. Sticker, V. Charwat, C. Kasper, G. Lepperdinger, Trends Biotechnol. 2014, 32, 245;
Z. Liu, X. Han, L. Qin, Adv. Healthcare Mater. 2016, 5, 871.
S. G. Uzel, O. C. Amadi, T. M. Pearl, R. T. Lee, P. T. So, R. D. Kamm, Small 2016, 12, 612;
Y. C. Chen, X. Lou, Z. Zhang, P. Ingram, E. Yoon, Sci. Rep. 2015, 5, 12175;
G. Bergstrom, J. Christoffersson, K. Schwanke, R. Zweigerdt, C. F. Mandenius, Lab Chip 2015, 15, 3242.
L. G. Villa-Diaz, Y. S. Torisawa, T. Uchida, J. Ding, N. C. Nogueira-de-Souza, K. S. O'Shea, S. Takayama, G. D. Smith, Lab Chip 2009, 9, 1749;
K. Kamei, M. Ohashi, E. Gschweng, Q. Ho, J. Suh, J. Tang, Z. T. For Yu, A. T. Clark, A. D. Pyle, M. A. Teitell, K. B. Lee, O. N. Witte, H. R. Tseng, Lab Chip 2010, 10, 1113;
S. Giulitti, E. Magrofuoco, L. Prevedello, N. Elvassore, Lab Chip 2013, 13, 4430.
G. G. Giobbe, F. Michielin, C. Luni, S. Giulitti, S. Martewicz, S. Dupont, A. Floreani, N. Elvassore, Nat. Methods 2015, 12, 637.
C. Luni, S. Giulitti, E. Serena, L. Ferrari, A. Zambon, O. Gagliano, G. G. Giobbe, F. Michielin, S. Knobel, A. Bosio, N. Elvassore, Nat. Methods 2016, 13, 446.
D. Park, J. Lim, J. Y. Park, S. H. Lee, Stem Cells Transl. Med. 2015, 4, 1352.
K. Kamei, Y. Mashimo, Y. Koyama, C. Fockenberg, M. Nakashima, M. Nakajima, J. Li, Y. Chen, Biomed. Microdev. 2015, 17, 36.
K. Kamei, Y. Hirai, M. Yoshioka, Y. Makino, Q. Yuan, M. Nakajima, Y. Chen, O. Tabata, Adv. Healthcare Mater. 2013, 2, 287.
H. Yoshioka, M. Mikami, Y. Mori, E. Tsuchida, J. Macromol. Sci. Pure Appl. Chem. 1994, A31, 121;
H. Yoshioka, M. Mikami, Y. Mori, E. Tsuchida, J. Macromol. Sci. Pure Appl. Chem. 1994, A31, 113.
T. Ludwig, V. Bergendahl, M. Levenstein, J. Yu, M. D. Probasco, J. Thomson, Nat. Methods 2006, 3, 637.
K. Watanabe, M. Ueno, D. Kamiya, A. Nishiyama, M. Matsumura, T. Wataya, J. B. Takahashi, S. Nishikawa, K. Muguruma, Y. Sasai, Nat. Biotechnol. 2007, 25, 681.
R. Krawetz, J. T. Taiani, S. Liu, G. Meng, X. Li, M. S. Kallos, D. E. Rancourt, Tissue Eng. C 2010, 16, 573.
H. Kempf, R. Olmer, C. Kropp, M. Ruckert, M. Jara-Avaca, D. Robles-Diaz, A. Franke, D. A. Elliott, D. Wojciechowski, M. Fischer, A. Roa Lara, G. Kensah, I. Gruh, A. Haverich, U. Martin, R. Zweigerdt, Stem Cell Rep. 2014, 3, 1132.
E. Tirotta, L. A. Kirby, M. N. Hatch, T. E. Lane, Stem Cell Res. 2012, 9, 208.
M. W. van der Helm, A. D. van der Meer, J. C. Eijkel, A. van den Berg, L. I. Segerink, Tissue Barriers 2016, 4, e1142493;
D. Bavli, S. Prill, E. Ezra, G. Levy, M. Cohen, M. Vinken, J. Vanfleteren, M. Jaeger, Y. Nahmias, Proc. Natl. Acad. Sci. USA 2016, 113, E2231;
W. Zhang, Y. S. Zhang, S. M. Bakht, J. Aleman, S. R. Shin, K. Yue, M. Sica, J. Ribas, M. Duchamp, J. Ju, R. B. Sadeghian, D. Kim, M. R. Dokmeci, A. Atala, A. Khademhosseini, Lab Chip 2016, 16, 1579;
B. Zhang, M. Montgomery, M. D. Chamberlain, S. Ogawa, A. Korolj, A. Pahnke, L. A. Wells, S. Masse, J. Kim, L. Reis, A. Momen, S. S. Nunes, A. R. Wheeler, K. Nanthakumar, G. Keller, M. V. Sefton, M. Radisic, Nat. Mater. 2016, 15, 669;
F. Zheng, F. Fu, Y. Cheng, C. Wang, Y. Zhao, Z. Gu, Small 2016, 12, 2253;
C. Oleaga, C. Bernabini, A. S. Smith, B. Srinivasan, M. Jackson, W. McLamb, V. Platt, R. Bridges, Y. Cai, N. Santhanam, B. Berry, S. Najjar, N. Akanda, X. Guo, C. Martin, G. Ekman, M. B. Esch, J. Langer, G. Ouedraogo, J. Cotovio, L. Breton, M. L. Shuler, J. J. Hickman, Sci. Rep. 2016, 6, 20030;
F. An, Y. Qu, Y. Luo, N. Fang, Y. Liu, Z. Gao, W. Zhao, B. Lin, Sci. Rep. 2016, 6, 25022;
S. N. Bhatia, D. E. Ingber, Nat. Biotechnol. 2014, 32, 760.
H. E. Abaci, M. L. Shuler, Integr. Biol. 2015, 7, 383;
P. G. Miller, M. L. Shuler, Biotechnol. Bioeng. 2016, 113, 2213;
S. H. Lee, S. K. Ha, I. Choi, N. Choi, T. H. Park, J. H. Sung, Biotechnol. J. 2016, 11, 746;
M. B. Esch, G. J. Mahler, T. Stokol, M. L. Shuler, Lab Chip 2014, 14, 3081.
C. A. Wells, R. Mosbergen, O. Korn, J. Choi, N. Seidenman, N. A. Matigian, A. M. Vitale, J. Shepherd, Stem Cell Res. 2013, 10, 387.
K. A. Le Cao, F. Rohart, L. McHugh, O. Korn, C. A. Wells, Genomics 2014, 103, 239.
G. K. Smyth, Stat. Appl. Genet. Mol. Biol. 2004, 3, Article 3;
G. K. Smyth, Y. H. Yang, T. Speed, Methods Mol. Biol. 2003, 224, 111.
M. D. Wilkerson, D. N. Hayes, Bioinformatics 2010, 26, 1572.
W. Huang da, B. T. Sherman, R. A. Lempicki, Nat. Protoc. 2009, 4, 44;
W. Huang da, B. T. Sherman, R. A. Lempicki, Nucl. Acids Res. 2009, 37, 1.