title: Cell Line Macroarray: An Alternative High-Throughput Platform to Analyze hiPSC Lines.
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.
authors: Institute of Genetic and Biomedical Research, National Research Council (IRGB-CNR), Milan, Italy (ALS, SB, IB).
authors: Alberto
authors: A
authors: La Spada
authors: Institute of Genetic and Biomedical Research, National Research Council (IRGB-CNR), Milan, Italy (ALS, SB, IB).
authors: Simona
authors: S
authors: Baronchelli
authors: Institute of Experimental Neurology (INSpe), Division of Neuroscience, San Raffaele Scientific Institute, Milan, Italy (LO, FR, GM).
authors: Linda
authors: L
authors: Ottoboni
authors: Institute of Experimental Neurology (INSpe), Division of Neuroscience, San Raffaele Scientific Institute, Milan, Italy (LO, FR, GM).
authors: Francesca
authors: F
authors: Ruffini
authors: Institute of Experimental Neurology (INSpe), Division of Neuroscience, San Raffaele Scientific Institute, Milan, Italy (LO, FR, GM).
authors: Gianvito
authors: G
authors: Martino
authors: IRCCS MultiMedica, Milan, Italy (NC, IB).
authors: Nunzia
authors: N
authors: Convertino
authors: Integrated Systems Engineering Srl, Milan, Italy (AN, ADB).
authors: Aikaterini
authors: A
authors: Ntai
authors: TissueGnostics GmbH, Vienna, Austria (TS)
authors: Tobias
authors: T
authors: Steiner
authors: Institute of Genetic and Biomedical Research, National Research Council (IRGB-CNR), Milan, Italy (ALS, SB, IB).|IRCCS MultiMedica, Milan, Italy (NC, IB).
authors: Ida
authors: I
authors: Biunno
authors: Integrated Systems Engineering Srl, Milan, Italy (AN, ADB).
authors: Andrea
authors: A
authors: De Blasio
mesh_terms: D002454:Cell Differentiation; D002460:Cell Line; D057166:High-Throughput Screening Assays; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D009474:Neurons / Q000166:cytology; D046888:Tissue Array Analysis / Q000379:methods*
keywords: cell line macroarray; hiPSC clones; high-throughput screening and quantification