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JRM Vol.25 No.4 pp. 623-630
doi: 10.20965/jrm.2013.p0623
(2013)

Paper:

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A High-Throughput Device for Patterned Differentiation of Embryoid Bodies

Xiaoming He*,**, Hiroshi Kimura*,**,***, and Teruo Fujii*,**

*Institute of Industrial Science, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan

**JST CREST, Tokyo, Japan

***Department of Mechanical Engineering, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan

Received:
March 9, 2013
Accepted:
May 21, 2013
Published:
August 20, 2013
Creative Commons License
Keywords:
iPS cells, embryoid body (EB), highthroughput analysis, microfluidic device, patterned differentiation
Abstract
Although ontogenesis in vivo may proceed in a spatiotemporally heterogeneous environment, in vitro differentiation of an embryoid body (EB) has been carried out in uniform conditions using conventional culture methods at low throughput. In the present study, a microfluidic device with multiple culture chambers for simultaneous patterned differentiation of multiple EBs of pluripotent stem cells is newly developed. Theoretical simulation and experiments using a suspension of fluorescent particles or fluorescent solution show that proper chemical gradients can be formed with almost no flow in the chambers. After multiple EBs are introduced into the device, these EBs move along the flow channel and into trapping cups. The EBs are pushed by air bubbles into the culture chambers. These multiple EBs can be cultured within the culture chambers after flowing culture medium removes the air bubble from the device. In our experiment, differentiation and proliferation of these multiple EBs are studied by exposing them to two different media for 6 days: one to induce differentiation and the other to keep the pluripotent and self-renewing state of the cells. It is shown that patterned differentiation of the multiple EBs is successfully conducted simultaneously in the device when these two media are perfused into the device. The results suggest that differentiation and proliferation of multiple EBs can be analyzed by applying chemical gradients in the present microfluidic device. This will be a helpful tool in a wide variety of experiments involving EBs or spheroids.
Cite this article as:
X. He, H. Kimura, and T. Fujii, “A High-Throughput Device for Patterned Differentiation of Embryoid Bodies,” J. Robot. Mechatron., Vol.25 No.4, pp. 623-630, 2013.
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References
  1. [1] K.Matsuura et al., “Creation of mouse embryonic stem cell-derived cardiac cell sheets,” Biomaterials, Vol.32, pp. 7355-7362, 2011.
  2. [2] H.-W. Wu, C.-C. Lin, and G.-B. Lee, “Stem cells in microfluidics,” Biomicrofluidics, Vol.5, 013401, 2011.
  3. [3] M. F. Pera and P. P. L. Tam, “Extrinsic regulation of pluripotent stem cells,” Nature, Vol.465, pp. 713-720, 2010.
  4. [4] J. El-Ali, P. K. Sorger, and K. F. Jensen, “Cells on chips,” Nature, Vol.442, pp. 403-411, 2006.
  5. [5] I. Meyvantsson and D. J. Beebe, “Cell Culture Models in Microfluidic Systems,” Annual Review of Analytic Chemistry, Vol.1, pp. 423-449, 2008.
  6. [6] J. Kawada, H. Kimura, H. Akutsu, Y. Sakai, and T. Fujii, “Spatiotemporally controlled delivery of soluble factors for stem cell differentiation,” Lab on a Chip, Vol.12, pp. 4508-4515, 2012.
  7. [7] W.-T. Fung, A. Beyzavi, P. Abgrall, N.-T. Nguyen, and H.-Y. Li, “Microfluidic platform for controlling the differentiation of embryoid bodies,” Lab on a Chip, Vol.9, pp. 2591-2595, 2009.
  8. [8] J. M. Karp et al., “Controlling size, shape and homogeneity of embryoid bodies using poly (ethylene glycol) microwells,” Lab on a Chip, Vol.7, pp. 786-794, 2007.
  9. [9] J. Park et al., “Microfabrication-based modulation of embryonic stem cell differentiation,” Lab on a Chip, Vol.7, pp. 1018-1028, 2007.
  10. [10] Y. Torisawa et al., “Efficient formation of uniform-sized embryoid bodies using a compartmentalized microchannel device,” Lab on a Chip, Vol.7, pp. 770-776, 2007.
  11. [11] E. Kang, Y. Y. Choi, Y. Jun, B. G. Chung, and S.-H. Lee, “Development of a multi-layer microfluidic array chip to culture and replate uniform-sized embryoid bodies without manual cell retrieval,” Lab on a Chip, Vol.10, pp. 2651-2654, 2010.
  12. [12] M. S. Kim et al., “Microfabricated embryonic stem cell divider for large-scale propagation of human embryonic stem cells,” Lab on a Chip, Vol.7, pp. 513-515, 2007.
  13. [13] C. Lochovsky, S. Yasotharan, and A. Gunther, “Bubbles no more: in-plane trapping and removal of bubbles in microfluidic devices,” Lab on a Chip, Vol.12, pp. 595-601, 2012.
  14. [14] F. Lugli and F. Zerbetto, “An introduction to bubble dynamics,” Physical chemistry Chemical Physics, Vol.9, pp. 2447-2456, 2007.
  15. [15] Y.-C. Toh and J. Voldman, “Fluid shear stress primes mouse embryonic stem cells for differentiation in a self-renewing environment via heparan sulfate proteoglycans transduction,” FASEB J., Vol.25, pp. 1208-1217, 2011.
  16. [16] K. Yamamoto et al., “Fluid shear stress induces differentiation of Flk-1-positive embryonic stem cells into vascular endothelial cells in vitro,” Am. J. Physiol. Heart Circ. Physiol., Vol.288, pp. 1915-1924, 2005.
  17. [17] E. Y. L. Fok and P. W. Zandstra, “Shear-Controlled Single-Step Mouse Embryonic Stem Cell Expansion and Embryoid Body–Based Differentiation,” Stem Cells, Vol.23, pp. 1333-1342, 2005.
  18. [18] W. Zheng, Z. Wang, W. Zhang, and X. Jiang, “A simple PDMSbased microfluidic channel design that removes bubbles for longterm on-chip culture of mammalian cells,” Lab on a chip, Vol.10, pp. 2906-2910, 2010.
  19. [19] S. A. Khan and S. Duraiswamy, “Controlling bubbles using bubbles-microfluidic synthesis of ultra-small gold nanocrystals with gas-evolving reducing agents,” Lab on a Chip, Vol.12, pp. 1807-1812, 2012.
  20. [20] A. R. Abate and D. A. Weitz, “Air-bubble-triggered drop formation in microfluidics,” Lab on a Chip, Vol.11, pp. 1713-1716, 2011.
  21. [21] A. P. Kotula and S. L. Anna, “Probing timescales for colloidal particle adsorption using slug bubbles in rectangular microchannels,” Soft Matter, 2012.

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