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

Authors :
La Spada A
Baronchelli S
Ottoboni L
Ruffini F
Martino G
Convertino N
Ntai A
Steiner T
Biunno I
De Blasio A
Source :
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society [J Histochem Cytochem] 2016 Dec; Vol. 64 (12), pp. 739-751. Date of Electronic Publication: 2016 Oct 26.
Publication Year :
2016

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.<br />Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.<br /> (© 2016 The Histochemical Society.)

Details

Language :
English
ISSN :
1551-5044
Volume :
64
Issue :
12
Database :
MEDLINE
Journal :
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
Publication Type :
Academic Journal
Accession number :
27798288
Full Text :
https://doi.org/10.1369/0022155416673969