Cell Line Macroarray: An Alternative High-Throughput Platform to Analyze hiPSC Lines
| Autor: | Ida Biunno, Francesca Ruffini, Linda Ottoboni, Andrea De Blasio, Tobias Steiner, Gianvito Martino, Aikaterini Ntai, Nunzia Convertino, Alberto La Spada, Simona Baronchelli |
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| Přispěvatelé: | La Spada, Alberto, Baronchelli, Simona, Ottoboni, Linda, Ruffini, Francesca, Martino, Gianvito, Convertino, Nunzia, Ntai, Aikaterini, Steiner, Tobia, Biunno, Ida, De Blasio, Andrea |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
High-Throughput Screening Assay Histology Cellular differentiation Cell Induced Pluripotent Stem Cells Computational biology Biology Proteomics Induced Pluripotent Stem Cell Cell Line 03 medical and health sciences medicine Humans high-throughput screening and quantification Induced pluripotent stem cell Neurons Cell growth cell line macroarray Cell Differentiation Articles Cell sorting Neuron Molecular biology High-Throughput Screening Assays 030104 developmental biology medicine.anatomical_structure Cell culture Tissue Array Analysis hiPSC clone Stem cell Anatomy Tissue Array Analysi Human |
| Zdroj: | The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society. 64(12) |
| ISSN: | 1551-5044 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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