Automated identification and location analysis of marked stem cells colonies in optical microscopy images

Vincenzo Paduano, Daniela Tagliaferri, Geppino Falco, Michele Ceccarelli

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Embryonic stem cells (ESCs) are characterized by two remarkable peculiarities: the capacity to propagate as undifferentiated cells (self-renewal) and the ability to differentiate in ectoderm, endoderm, and mesoderm derivatives (pluripotency). Although the majority of ESCs divide without losing the pluripotency, it has become evident that ESC cultures consists of multiple cell populations highlighted by the expression of early germ lineage markers during spontaneous differentiation. Hence, the identification and characterization of ESCs subpopulations represents an efficient approach to improve the comprehension of correlation between gene expression and cell specification status. To study markers of ESCs heterogeneity, we developed an analysis pipeline which can automatically process images of stem cell colonies in optical microscopy. The question we try to address is to find out the statistically significant preferred locations of the marked cells. We tested our algorithm on a set of images of stem cell colonies to analyze the expression pattern of the Zscan4 gene, which was an elite candidate gene to be studied because it is specifically expressed in subpopulation of ESCs. To validate the proposed method we analyzed the behavior of control genes whose pattern had been associated to biological status such as differentiation (EndoA), pluripotency (Pou5f1), and pluripotency fluctuation (Nanog). We found that Zscan4 is not uniformly expressed inside a stem cell colony, and that it tends to be expressed towards the center of the colony, moreover cells expressing Zscan4 cluster each other. This is of significant importance because it allows us to hypothesize a biological status where the cells expressing Zscan4 are preferably associated to the inner of colonies suggesting pluripotent cell status features, and the clustering between themselves suggests either a colony paracrine effect or an early phase of cell specification through proliferation. Also, the analysis on the control genes showed that they behave as expected.

Original languageEnglish
Article numbere80776
JournalPLoS One
Volume8
Issue number12
DOIs
Publication statusPublished - 9 Dec 2013
Externally publishedYes

Fingerprint

Stem cells
Optical microscopy
Embryonic Stem Cells
stem cells
Microscopy
microscopy
embryonic stem cells
Stem Cells
cells
Genes
Cells
genes
Endoderm
Ectoderm
Behavior Control
Aptitude
Gene Expression Profiling
Mesoderm
Specifications
Cluster Analysis

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Automated identification and location analysis of marked stem cells colonies in optical microscopy images. / Paduano, Vincenzo; Tagliaferri, Daniela; Falco, Geppino; Ceccarelli, Michele.

In: PLoS One, Vol. 8, No. 12, e80776, 09.12.2013.

Research output: Contribution to journalArticle

Paduano, Vincenzo ; Tagliaferri, Daniela ; Falco, Geppino ; Ceccarelli, Michele. / Automated identification and location analysis of marked stem cells colonies in optical microscopy images. In: PLoS One. 2013 ; Vol. 8, No. 12.
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