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A micro-channel-well system for culture and differentiation of embryonic stem cells on different types of substrate

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Abstract

We have developed a combined micro-channel and micro-well system for easy cell loading, culture and post-culture operation on a chip. To demonstrate the reliability of the system, on chip cell culture and differentiation were performed with different types of substrates made of culture dish, glass cover slide and polydimethylsiloaxe (PDMS). As expected, mouse embryo fibroblasts (MEF) showed different adhesion and growth rate on different substrates. When embryonic stem (ES) cells were co-cultured with MEFs, the formation of ES colonies is efficient on both glass and Petri dish, although PDMS could also be used. Finally, ES cell differentiation with neuron growth factors was performed on different substrates, showing clear advantages of using culture Petri dish over both glass and PDMS.

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Abbreviations

ES:

embryonic stem

EB:

embryo bodies

mES:

mouse embryonic stem

MEF:

mouse embryo fibroblasts

LIF:

leukemia inhibitory factor

PS:

polystyrene

RA:

retinoic acid

DAPI:

4′-6-Diamidino-2-phenylindole

DMEM:

Dulbecco’s Modified Eagle’s Medium

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Acknowledgments

The authors would like to thank Prof. K. Yoshikawa and Dr. Y. Ma of Kyoto University for supports and discussions. This work was partially supported by Grants-in-Aid for Scientific Research from the Japanese Government (No. 19684015, 20034016).

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Authors and Affiliations

  1. Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University, Kyoto, 606-8507, Japan

    Li Liu, Chunxiong Luo, Xiaofeng Ni, Li Wang, Kaori Yamauchi, Shinichi M. Nomura, Norio Nakatsuji & Yong Chen

  2. Center for Microfluidic and Nanotechnology, Peking University, Beijing, 100871, China

    Chunxiong Luo

  3. Ecole Normale Supérieure, CNRS-ENS-UPMC UMR 8640, 24 rue Lhomond, Paris, 75231, France

    Xiaofeng Ni, Li Wang & Yong Chen

  4. PRESTO, Japan Science and Technology Agency (JST), Tokyo, 102-0075, Japan

    Shinichi M. Nomura

Authors
  1. Li Liu
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  2. Chunxiong Luo
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  3. Xiaofeng Ni
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  4. Li Wang
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  5. Kaori Yamauchi
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  6. Shinichi M. Nomura
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  7. Norio Nakatsuji
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  8. Yong Chen
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Corresponding author

Correspondence to Yong Chen.

Additional information

Li Liu and Chunxiong Luo contributed equally to this work.

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Liu, L., Luo, C., Ni, X. et al. A micro-channel-well system for culture and differentiation of embryonic stem cells on different types of substrate. Biomed Microdevices 12, 505–511 (2010). https://doi.org/10.1007/s10544-010-9407-4

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  • DOI: https://doi.org/10.1007/s10544-010-9407-4

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