Abstract
Liver is composed of various kinds of cells, including hepatic parenchymal cells (hepatocytes) and nonparenchymal cells, and separation of these cells is essential for cellular therapies and pharmacological and metabolic studies. Here, we present microfluidic devices for purely hydrodynamic and size-dependent separation of liver cells, which utilize hydrodynamic filtration. By continuously introducing cell suspension into a microchannel with multiple side-branch channels, cells smaller than a specific size are removed from the mainstream, while large cells are focused onto a sidewall in the microchannel and then separated into two or three groups. Two types of PDMS-glass hybrid microdevices were fabricated, and rat liver cells were successfully separated. Also, cell size, morphology, viability and several cell functions were analyzed, and the separation performances of the microfluidic devices were compared to that of a conventional centrifugal technique. The results showed that the presented microfluidic devices are low-cost and suitable for clinical use, and capable of highly functional separation with relatively high-speed processing.
Similar content being viewed by others
References
H. Andersson, A. van den Berg, Sens. Actuators B 92, 315 (2003)
K. Asahina, M. Shiokawa, T. Ueki, C. Yamasaki, A. Aratani, C. Tateno, K. Yoshizato, Biochem. Biophys. Res. Commun. 342, 1160 (2006)
P.A. Auroux, D. Iossifidis, D.R. Reyes, A. Manz, Anal. Chem. 74, 2637 (2002)
D.C. Duffy, J.C. McDonald, O.J.A. Schueller, G.M. Whitesides, Anal. Chem. 70, 4974 (1998)
J. El-Ali, P.K. Sorger, K.F. Jensen, Nature 442, 403 (2006)
K. Hosokawa, K. Sato, N. Ichikawa, M. Maeda, Lab Chip 4, 181 (2004)
L.R. Huang, E.C. Cox, R.H. Austin, J.C. Sturm, Science 304, 987 (2004)
B.J. Kane, M.J. Zinner, M.L. Yarmush, M. Toner, Anal. Chem. 78, 4291 (2006)
S. Katayama, C. Tateno, T. Asahara, K. Yoshizato, Am. J. Pathol. 158, 97 (2001)
D.L. Knook, E.C. Sleyster, Exp. Cell Res. 99, 444 (1976)
D.L. Knook, A.M. Seffelaar, A.M. de Leeuw, Exp. Cell Res. 139, 468 (1982)
B.L. Kreamer, J.L. Staecker, N. Sawada, G.L. Sattler, M.T.S. Hsia, H.C. Pitot, In Vitro Cell. Dev. Biol. 22, 201 (1986)
T. Mitaka, M. Mikami, G.L. Sattler, H.C. Pitot, Y. Mochizuki, Hepatology 16, 440 (1992)
O. Morin, F. Goulet, C. Normand, in Cell Biology Reviews (Springer International, New York, 1988), pp. 1–69
L. Mossin, H. Blankson, H. Huitfeldt, P.O. Seglen, Exp. Cell Res. 214, 551 (1994)
S. K. Murthy, P. Sethu, G. Vunjak-Novakovic, M. Toner, M. Radisic, Biomed. Microdev. 8, 231 (2006)
K.H. Nam, W.J. Chang, H. Hong, S.M. Lim, D.I. Kim, Y.M. Koo, Biomed. Microdev. 7, 189 (2005)
M. Nilsson, T. Berg, Biochim. Biophys. Acta 497, 171 (1977)
K. Ohashi, F. Park, M.A. Kay, J. Mol. Med. 79, 617 (2001)
P.O. Seglen, Methods Cell Biol. 13, 29 (1976)
P. Sethu, A. Sin, M. Toner, Lab Chip 6, 83 (2006)
S.S. Shevkoplyas, T. Yoshida, L.L. Munn, M.W. Bitensky, Anal. Chem. 77, 933 (2005)
S.H. Sigal, S. Gupta, D.F. Gebhard Jr., P. Holst, D. Neufeld, L.M. Reid, Differentiation 59, 35 (1995)
B. Singh, B. Borrebaek, H. Osmundsen, Acta Physiol. Scand. 117, 497 (1983)
J. Takagi, M. Yamada, M. Yasuda, M. Seki, Lab Chip 5, 778 (2005)
C. Tateno, K. Yoshizato, Am. J. Pathol. 149, 1593 (1996)
Y. Tsuda, A. Kikuchi, M. Yamato, G.P. Chen, T. Okano, Biochem. Biophys. Res. Commun. 348, 937 (2006)
M. Yamada, M. Seki, Lab Chip 5, 1233 (2005)
M. Yamada, M. Seki, Anal. Chem. 78, 1357 (2006)
M. Yamada, V. Kasim, M. Nakashima, J. Edahiro, M. Seki, Biotechnol. Bioeng. 88, 489 (2004)
Acknowledgements
This research was supported in part by the Core Research for Evolution Science and Technology from the Japan Science and Technology Agency, and by Grant-in-aid for JSPS Fellowship. We are grateful to Ms. Chinatsu Kohno for her technical assistance.
Author information
Authors and Affiliations
Corresponding author
Electronic Supplementary Material
Below is the link to the supplementary material
Rights and permissions
About this article
Cite this article
Yamada, M., Kano, K., Tsuda, Y. et al. Microfluidic devices for size-dependent separation of liver cells. Biomed Microdevices 9, 637–645 (2007). https://doi.org/10.1007/s10544-007-9055-5
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10544-007-9055-5