Cell Line Macroarray: An Alternative High-Throughput Platform to Analyze hiPSC Lines

Alberto La Spada, Simona Baronchelli, Linda Ottoboni, Francesca Ruffini, Gianvito Martino, Nunzia Convertino, Aikaterini Ntai, Tobias Steiner, Ida Biunno, Andrea De Blasio

Research output: Contribution to journalArticle

Abstract

In the past decade, tissue microarray (TMA) technology has evolved as an innovative tool for high-throughput proteomics analysis and mainly for biomarker validation. Similarly, enormous amount of data can be obtained from the cell line macroarray (CLMA) technology, which developed from the TMA using formalin-fixed, paraffin-embedded cell pellets. Here, we applied CLMA technology in stem cell research and in particular to identify bona fide neogenerated human induced pluripotent stem cell (hiPSC) clones suitable for down the line differentiation. All hiPSC protocols generate tens of clones, which need to be tested to determine genetically stable cell lines suitable for differentiation. Screening methods generally rely on fluorescence-activated cell sorting isolation and coverslip cell growth followed by immunofluorescence; these techniques could be cumbersome. Here, we show the application of CLMA to identify neogenerated pluripotent cell colonies and neuronal differentiated cell products. We also propose the use of the automated image analyzer, TissueQuest, as a reliable tool to quickly select the best clones, based upon the level of expression of multiple pluripotent biomarkers.

Original languageEnglish
Pages (from-to)739-751
Number of pages13
JournalJournal of Histochemistry and Cytochemistry
Volume64
Issue number12
DOIs
Publication statusPublished - 2016

Fingerprint

Induced Pluripotent Stem Cells
Cell Line
Clone Cells
Technology
Biomarkers
Stem Cell Research
Cell Separation
Paraffin
Proteomics
Formaldehyde
Fluorescent Antibody Technique
Flow Cytometry
Growth

Keywords

  • cell line macroarray
  • high-throughput screening and quantification
  • hiPSC clones

ASJC Scopus subject areas

  • Anatomy
  • Histology

Cite this

La Spada, A., Baronchelli, S., Ottoboni, L., Ruffini, F., Martino, G., Convertino, N., ... De Blasio, A. (2016). Cell Line Macroarray: An Alternative High-Throughput Platform to Analyze hiPSC Lines. Journal of Histochemistry and Cytochemistry, 64(12), 739-751. https://doi.org/10.1369/0022155416673969

Cell Line Macroarray : An Alternative High-Throughput Platform to Analyze hiPSC Lines. / La Spada, Alberto; Baronchelli, Simona; Ottoboni, Linda; Ruffini, Francesca; Martino, Gianvito; Convertino, Nunzia; Ntai, Aikaterini; Steiner, Tobias; Biunno, Ida; De Blasio, Andrea.

In: Journal of Histochemistry and Cytochemistry, Vol. 64, No. 12, 2016, p. 739-751.

Research output: Contribution to journalArticle

La Spada, A, Baronchelli, S, Ottoboni, L, Ruffini, F, Martino, G, Convertino, N, Ntai, A, Steiner, T, Biunno, I & De Blasio, A 2016, 'Cell Line Macroarray: An Alternative High-Throughput Platform to Analyze hiPSC Lines', Journal of Histochemistry and Cytochemistry, vol. 64, no. 12, pp. 739-751. https://doi.org/10.1369/0022155416673969
La Spada, Alberto ; Baronchelli, Simona ; Ottoboni, Linda ; Ruffini, Francesca ; Martino, Gianvito ; Convertino, Nunzia ; Ntai, Aikaterini ; Steiner, Tobias ; Biunno, Ida ; De Blasio, Andrea. / Cell Line Macroarray : An Alternative High-Throughput Platform to Analyze hiPSC Lines. In: Journal of Histochemistry and Cytochemistry. 2016 ; Vol. 64, No. 12. pp. 739-751.
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