Automated Measurement of Cobblestone Morphology for Characterizing Stem Cell Derived Retinal Pigment Epithelial Cell Cultures

J Ocul Pharmacol Ther. 2016 Jun;32(5):331-9. doi: 10.1089/jop.2015.0163. Epub 2016 May 18.


Purpose: Assessing the morphologic properties of cells in microscopy images is an important task to evaluate cell health, identity, and purity. Typically, subjective visual assessments are accomplished by an experienced researcher. This subjective human step makes transfer of the evaluation process from the laboratory to the cell manufacturing facility difficult and time consuming.

Methods: Automated image analysis can provide rapid, objective measurements of cultured cells, greatly aiding manufacturing, regulatory, and research goals. Automated algorithms for classifying images based on appearance characteristics typically either extract features from the image and use those features for classification or use the images directly as input to the classification algorithm. In this study we have developed both feature and nonfeature extraction methods for automatically measuring "cobblestone" structure in human retinal pigment epithelial (RPE) cell cultures.

Results: A new approach using image compression combined with a Kolmogorov complexity-based distance metric enables robust classification of microscopy images of RPE cell cultures. The automated measurements corroborate determinations made by experienced cell biologists. We have also developed an approach for using steerable wavelet filters for extracting features to characterize the individual cellular junctions.

Conclusions: Two image analysis techniques enable robust and accurate characterization of the cobblestone morphology that is indicative of viable RPE cultures for therapeutic applications.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Algorithms
  • Automation
  • Cell Culture Techniques*
  • Cell Shape*
  • Cells, Cultured
  • Epithelial-Mesenchymal Transition
  • Humans
  • Image Interpretation, Computer-Assisted / methods*
  • Pattern Recognition, Automated / methods
  • Retinal Pigment Epithelium / cytology*
  • Stem Cells / cytology*