Jump to main content
Jump to site search
Publishing
Advanced

Issue 12, 2009
Previous Article Next Article

Microfluidic culture of single human embryonic stem cell colonies

Luis Gerardo Villa-Diaz,a  Yu-suke Torisawa,b  Tomoyuki Uchida,b  Jun Ding,a  Naiara Correa Nogueira-de-Souza,a  Kathy Sue O'Shea,c  Shuichi Takayama*bd  and  Gary Daniel Smith*aefg 
Author affiliations

* Corresponding authors

a Departments of Obstetrics and Gynecology, University of Michigan, Ann Arbor, USA
E-mail: smithgd@umich.edu

b Departments of Biomedical Engineering, University of Michigan, Ann Arbor, USA
E-mail: takayama@umich.edu

c Departments of Cell and Developmental Biology, University of Michigan, Ann Arbor, USA

d Departments of Macromolecular Science and Engineering, University of Michigan, Ann Arbor, USA

e Departments of Urology, University of Michigan, Ann Arbor, USA

f Departments of Molecular and Integrated Physiology, University of Michigan, Ann Arbor, USA

g Departments of Reproductive Sciences Program, University of Michigan, Ann Arbor, USA

Abstract

We have developed a miniaturized microfluidic culture system that allows experimentation on individual human embryonic stem cell (hESC) colonies in dynamic (flow applied) or static (without flow) conditions. The system consists of three inlet channels that converge into a cell-culture channel and provides the capability to spatially and temporally deliver specific treatments by using patterned laminar fluid flow to different parts of a single hESC colony. We show that microfluidic culture for 96 h with or without flow results in similar maintenance of hESC self-renewal, the capability to differentiate into three germ cell lineages, and to maintain a normal karyotype, as in standard culture dishes. Localized delivery of a fluorescent nucleic acid dye was achieved with laminar flow, producing staining only in nuclei of exposed cells. Likewise, cells in desired regions of colonies could be removed with enzymatic treatment and collected for analysis. Re-coating the enzyme treated area of the channel with extracellular matrix led to re-growth of hESC colonies into this region. Our study demonstrates the culture of hESCs in a microfluidic device that can deliver specific treatments to desired regions of a single colony. This miniaturized culture system allows in situ treatment and analysis with the ability to obtain cell samples from part of a colony without micromanipulation and to perform sensitive molecular analysis while permitting further growth of the hESC colony.

Graphical abstract: Microfluidic culture of single human embryonic stem cell colonies

Back to tab navigation
Download options Please wait...

Publication details

The article was received on 14 Nov 2008, accepted on 02 Mar 2009 and first published on 24 Mar 2009


Article type: Paper
DOI: 10.1039/B820380F
Citation: Lab Chip, 2009,9, 1749-1755
  •   Request permissions

    Microfluidic culture of single human embryonic stem cell colonies

    L. G. Villa-Diaz, Y. Torisawa, T. Uchida, J. Ding, N. C. Nogueira-de-Souza, K. S. O'Shea, S. Takayama and G. D. Smith, Lab Chip, 2009, 9, 1749
    DOI: 10.1039/B820380F

    If you are not the author of this article and you wish to reproduce material from it in a third party non-RSC publication you must formally request permission using RightsLink. Go to our Instructions for using RightsLink page for details.

    Authors contributing to RSC publications (journal articles, books or book chapters) do not need to formally request permission to reproduce material contained in this article provided that the correct acknowledgement is given with the reproduced material.

    Reproduced material should be attributed as follows:

    • For reproduction of material from NJC:
      Reproduced from Ref. XX with permission from the Centre National de la Recherche Scientifique (CNRS) and The Royal Society of Chemistry.
    • For reproduction of material from PCCP:
      Reproduced from Ref. XX with permission from the PCCP Owner Societies.
    • For reproduction of material from PPS:
      Reproduced from Ref. XX with permission from the European Society for Photobiology, the European Photochemistry Association, and The Royal Society of Chemistry.
    • For reproduction of material from all other RSC journals and books:
      Reproduced from Ref. XX with permission from The Royal Society of Chemistry.

    If the material has been adapted instead of reproduced from the original RSC publication "Reproduced from" can be substituted with "Adapted from".

    In all cases the Ref. XX is the XXth reference in the list of references.

    If you are the author of this article you do not need to formally request permission to reproduce figures, diagrams etc. contained in this article in third party publications or in a thesis or dissertation provided that the correct acknowledgement is given with the reproduced material.

    Reproduced material should be attributed as follows:

    • For reproduction of material from NJC:
      [Original citation] - Reproduced by permission of The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS) and the RSC
    • For reproduction of material from PCCP:
      [Original citation] - Reproduced by permission of the PCCP Owner Societies
    • For reproduction of material from PPS:
      [Original citation] - Reproduced by permission of The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC
    • For reproduction of material from all other RSC journals:
      [Original citation] - Reproduced by permission of The Royal Society of Chemistry

    If you are the author of this article you still need to obtain permission to reproduce the whole article in a third party publication with the exception of reproduction of the whole article in a thesis or dissertation.

    Information about reproducing material from RSC articles with different licences is available on our Permission Requests page.

Search articles by author

Spotlight

Advertisements