High-Throughput and Cost-Effective Characterization of Induced Pluripotent Stem Cells
| Autor: | Angelo Arias, Agnieszka D'Antonio-Chronowska, Cheryl Herrera, Jason L. Nathanson, Hiroko Matsui, Grace Woodruff, Matteo D’Antonio, Kelly A. Frazer, Lawrence S.B. Goldstein, Athanasia D. Panopoulos, Gene W. Yeo, Sol M. Reyna, Roy Williams |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Somatic cell Cost-Benefit Analysis Cellular differentiation Regenerative Medicine Bioinformatics Biochemistry fluorescent cell barcoding 2.1 Biological and endogenous factors Myocytes Cardiac Stem Cell Research - Induced Pluripotent Stem Cell - Non-Human Aetiology Induced pluripotent stem cell lcsh:QH301-705.5 Throughput (business) pluripotency characterization Neurons lcsh:R5-920 Stem Cell Research - Induced Pluripotent Stem Cell - Human medicine.diagnostic_test Cell Differentiation Cellular Reprogramming Phenotype digital karyotyping 3. Good health qPCR lcsh:Medicine (General) Cardiac Reprogramming Biotechnology Resource Genotype induced pluripotent stem cells Clinical Sciences Computational biology Biology Cell Line high-throughput methods Flow cytometry 03 medical and health sciences Genetics medicine Humans Myocytes SNP arrays Stem Cell Research - Induced Pluripotent Stem Cell flow cytometry Genetic Variation Cell Biology Stem Cell Research High-Throughput Screening Assays differentiation potential 030104 developmental biology lcsh:Biology (General) Cell culture Karyotyping Generic health relevance Biochemistry and Cell Biology Biomarkers Developmental Biology |
| Zdroj: | Stem cell reports, vol 8, iss 4 Stem Cell Reports, Vol 8, Iss 4, Pp 1101-1111 (2017) Stem Cell Reports |
| ISSN: | 2213-6711 |
| Popis: | Summary Reprogramming somatic cells to induced pluripotent stem cells (iPSCs) offers the possibility of studying the molecular mechanisms underlying human diseases in cell types difficult to extract from living patients, such as neurons and cardiomyocytes. To date, studies have been published that use small panels of iPSC-derived cell lines to study monogenic diseases. However, to study complex diseases, where the genetic variation underlying the disorder is unknown, a sizable number of patient-specific iPSC lines and controls need to be generated. Currently the methods for deriving and characterizing iPSCs are time consuming, expensive, and, in some cases, descriptive but not quantitative. Here we set out to develop a set of simple methods that reduce cost and increase throughput in the characterization of iPSC lines. Specifically, we outline methods for high-throughput quantification of surface markers, gene expression analysis of in vitro differentiation potential, and evaluation of karyotype with markedly reduced cost. Graphical Abstract Highlights • Combining three high-throughput methods provides low-cost characterization of iPSCs • iPSC line heterogeneity is assessed by fluorescent cell barcoding flow cytometry • 12-gene qPCR enables gene expression analysis of in vitro differentiation potential • SNP arrays provide inexpensive high-resolution digital karyotyping Working as part of the NHLBI NextGen consortium, D'Antonio and colleagues developed three simple methods that reduce cost and increase throughput in the characterization of iPSCs. These methods include: (1) fluorescent cell barcoding flow cytometry to investigate heterogeneity; (2) gene expression analysis to examine in vitro differentiation potential; and (3) high-resolution digital karyotyping to detect chromosomal aberrations. |
| Databáze: | OpenAIRE |
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