Cell Line Macroarray: An Alternative High-Throughput Platform to Analyze hiPSC Lines. (December 2016)

• Record Type: Journal Article
• Authors: La Spada, Alberto; Baronchelli, Simona; Ottoboni, Linda; Ruffini, Francesca; Martino, Gianvito; Convertino, Nunzia; Ntai, Aikaterini; Steiner, Tobias; Biunno, Ida; De Blasio, Andrea
• Is Part Of: Journal of histochemistry and cytochemistry. Volume 64:Number 12(2016:Dec.)
• Page Start: 739
• Page End: 751
• Publication Date: 2016-12
• Subjects: cell line macroarray -- high-throughput screening and quantification -- hiPSC clones; Histochemistry -- Periodicals; Cytochemistry -- Periodicals; Histocytochemistry -- methods; Histological Techniques; 572.05
• DOI: 10.1369/0022155416673969
• Languages: English
• ISSNs: 0022-1554

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.