Development of a new approach to aid in visual identification of murine iPS colonies using a fuzzy logic decision support system

PLoS One. 2013 Aug 8;8(8):e70605. doi: 10.1371/journal.pone.0070605. eCollection 2013.

Abstract

The a priori identification of induced pluripotent stem cells remains a challenge. Being able to quickly identify the most embryonic stem cell-similar induced pluripotent stem cells when validating results could help to reduce costs and save time. In this context, tools based on non-classic logic can be useful in creating aid-systems based on visual criteria. True colonies when viewed at 100x magnification have been found to have the following 3 characteristics: a high degree of border delineation, a more uniform texture, and the absence of a cracked texture. These visual criteria were used for fuzzy logic modeling. We investigated the possibility of predicting the presence of alkaline phosphatase activity, typical of true induced pluripotent stem cell colonies, after 25 individuals, with varying degrees of experience in working with murine iPS cells, categorized the images of 136 colonies based on visual criteria. Intriguingly, the performance evaluation by area under the ROC curve (16 individuals with satisfactory performance), Spearman correlation (all statistically significant), and Cohen's Kappa agreement analysis (all statistically significant) demonstrates that the discriminatory capacity of different evaluators are similar, even those who have never cultivated cells. Thus, we report on a new system to facilitate visual identification of murine- induced pluripotent stem cell colonies that can be useful for staff training and opens the possibility of exploring visual characteristics of induced pluripotent stem cell colonies with their functional peculiarities. The fuzzy model has been integrated as a web-based tool named "2see-iPS" which is freely accessed at http://genetica.incor.usp.br/2seeips/.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Animals
  • Cells, Cultured
  • Fuzzy Logic*
  • Image Processing, Computer-Assisted / methods*
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Mice

Substances

  • Alkaline Phosphatase

Grants and funding

VB and CBS were recipients of training fellowships from Fundação de Amparo à Pesquisa do Estado de São Paulo - FAPESP (2008/52334-1, 2008/52335-8, respectively). VB was enrolled in the PhD program at FMUSP. This work was funded by grants from FAPESP (2008/52436-9), CNPq (573887/2008-0) and an unrestricted gift from Roche Pharmaceutical. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.