Reichen, M;
Macown, RJ;
Jaccard, N;
Super, A;
Ruban, L;
Griffin, LD;
Veraitch, FS;
(2012)
Microfabricated modular scale-down device for regenerative medicine process development.
PLOS One
, 7
(12)
, Article e52246. 10.1371/journal.pone.0052246.
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Abstract
The capacity of milli and micro litre bioreactors to accelerate process development has been successfully demonstrated in traditional biotechnology. However, for regenerative medicine present smaller scale culture methods cannot cope with the wide range of processing variables that need to be evaluated. Existing microfabricated culture devices, which could test different culture variables with a minimum amount of resources (e.g. expensive culture medium), are typically not designed with process development in mind. We present a novel, autoclavable, and microfabricated scale-down device designed for regenerative medicine process development. The microfabricated device contains a re-sealable culture chamber that facilitates use of standard culture protocols, creating a link with traditional small-scale culture devices for validation and scale-up studies. Further, the modular design can easily accommodate investigation of different culture substrate/extra-cellular matrix combinations. Inactivated mouse embryonic fibroblasts (iMEF) and human embryonic stem cell (hESC) colonies were successfully seeded on gelatine-coated tissue culture polystyrene (TC-PS) using standard static seeding protocols. The microfluidic chip included in the device offers precise and accurate control over the culture medium flow rate and resulting shear stresses in the device. Cells were cultured for two days with media perfused at 300 µl.h(-1) resulting in a modelled shear stress of 1.1×10(-4) Pa. Following perfusion, hESC colonies stained positively for different pluripotency markers and retained an undifferentiated morphology. An image processing algorithm was developed which permits quantification of co-cultured colony-forming cells from phase contrast microscope images. hESC colony sizes were quantified against the background of the feeder cells (iMEF) in less than 45 seconds for high-resolution images, which will permit real-time monitoring of culture progress in future experiments. The presented device is a first step to harness the advantages of microfluidics for regenerative medicine process development.
| Type: | Article |
|---|---|
| Title: | Microfabricated modular scale-down device for regenerative medicine process development. |
| Location: | United States |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1371/journal.pone.0052246 |
| Publisher version: | http://dx.doi.org/10.1371/journal.pone.0052246 |
| Language: | English |
| Additional information: | © 2012 Reichen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The authors gratefully acknowledge the Engineering and Physical Sciences Research Council EPSRC (‘First Grant’, EP/I005471/1); the Department of Biochemical Engineering, UCL, for providing funding during Marcel Reichen's PhD studentship; the Peter Dunnill Scholarship, UCL’s Engineering Sciences Faculty and UCL’s Overseas Research Scholarship for the funding of Rhys J. Macown’s PhD studentship; the British Heart Foundation (BHF SP/08/004) and UCL’s Centre for Mathematics and Physics in the Life Sciences and Experimental Biology for funding Nicolas Jaccard’s PhD studentship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Computer Science |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1361548 |
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