Loughborough University
Browse

File(s) under permanent embargo

Reason: This item is currently closed access.

Automated image analysis with the potential for process quality control applications in stem cell maintenance and differentiation

journal contribution
posted on 2016-05-12, 13:50 authored by David Smith, Katie GlenKatie Glen, Rob ThomasRob Thomas
The translation of laboratory processes into scaled production systems suitable for manu-facture is a significant challenge for cell based therapies; in particular there is a lack ofanalytical methods that are informative and efficient for process control. Here the potentialof image analysis as one part of the solution to this issue is explored, using pluripotent stemcell colonies as a valuable and challenging exemplar. The Cell-IQ live cell imaging platformwas used to build image libraries of morphological culture attributes such as colony “edge,”“core periphery” or “core” cells. Conventional biomarkers, such as Oct3/4, Nanog, andSox-2, were shown to correspond to specific morphologies using immunostaining and flowcytometry techniques. Quantitative monitoring of these morphological attributes in-processusing the reference image libraries showed rapid sensitivity to changes induced by differentmedia exchange regimes or the addit ion of mesoderm lineage inducing cytokine BMP4. Theimaging sample size to precision relationship was defined for each morphological attributeto show that this sensitivity could be achieved with a relatively low imaging sample. Further,the morphological state of single colonies could be correlated to individual colony out-comes; smaller colonies were identified as optimum for homogenous early mesoderm differ-entiation, while larger colonies maintained a morphologically pluripotent core. Finally, weshow the potential of the same image libraries to assess cell number in culture with accu-racy comparable to sacrificial digestion and counting. The data supports a potentiallypowerful role for quantitative image analysis in the setting of in-process specifications, andalso for screening the effects of process actions during development, which is highly comple-mentary to current analysis in optimization and manufacture.

Funding

This work was supported by EPSRC Centre for Innovative Manufacturing grant EP/H028277/1. David Smith was funded by EPSRC Doctoral Training Centre in Regenerative Medicine grant EP/F500491/1 alongside CM-Technologies industry sponsor.

History

School

  • Mechanical, Electrical and Manufacturing Engineering

Published in

BIOTECHNOLOGY PROGRESS

Volume

32

Issue

1

Pages

215 - 223 (9)

Citation

SMITH, D., GLEN, K.E. and THOMAS, R.J., 2016. Automated image analysis with the potential for process quality control applications in stem cell maintenance and differentiation. Biotechnology Progress, 32 (1), pp. 215 - 223.

Publisher

American Institute of Chemical Engineers (Wiley)

Version

  • VoR (Version of Record)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at: https://creativecommons.org/licenses/by-nc-nd/4.0/

Publication date

2015-11-11

Notes

This article will be made open access as soon as we have the gold open access version of the paper.

ISSN

8756-7938

Language

  • en

Usage metrics

    Loughborough Publications

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC