Abstract
Induced pluripotent stem cells (iPSCs) derived from human somatic cells have created new opportunities to generate disease-relevant cells. Thus, as the use of patient-derived stem cells has become more widespread, having a workflow to monitor each line is critical. This ensures iPSCs pass a suite of quality-control measures, promoting reproducibility across experiments and between labs. With this in mind, we established a multistep workflow to assess our newly generated iPSCs. Our workflow tests four benchmarks: cell growth, genomic stability, pluripotency, and the ability to form the three germline layers. We also outline a simple test for assessing cell growth and highlight the need to compare different growth media. Genomic integrity in the human iPSCs is analyzed by G-band karyotyping and a qPCR-based test for the detection of common karyotypic abnormalities. Finally, we confirm that the iPSC lines can differentiate into a given cell type, using a trilineage assay, and later confirm that each iPSC can be differentiated into one cell type of interest, with a focus on the generation of cortical neurons. Taken together, we present a multistep quality-control workflow to evaluate newly generated iPSCs and detail the findings on these lines as they are tested within the workflow.
Highlights
Human pluripotent stem cells can give rise to almost any cell type when exposed to the appropriate developmental cues, holding enormous potential for tissue engineering, regenerative medicine, and disease modeling
We focused on cortical neurons, as methods to generate these neurons from Induced pluripotent stem cells (iPSCs) are well-established, and represent a cell type broadly studied across disease areas of the brain, from Alzheimer’s disease to neurodevelopmental disorders [16,17]
All iPSC lines attached onto Matrigel-coated plates and presented with similar morphology when maintained in either medium
Summary
Human pluripotent stem cells can give rise to almost any cell type when exposed to the appropriate developmental cues, holding enormous potential for tissue engineering, regenerative medicine, and disease modeling. The growing number of iPSC lines and National Institutes of Health (NIH)-registered human embryonic stem cell (ESC) lines ensures that patient-derived pluripotent stem cells are readily available to researchers, helping to accelerate our understanding of biology and disease and the development of therapies across disease areas [1]. These advances underscore the need for iPSC quality standards that are sufficiently stringent to ensure that findings can be compared and results reproduced across laboratories [2,3]. By assaying growth rate, genomic instability, pluripotency, and differentiation for a cell line, we can establish how well it performs and if it can used to generate the cells needed for downstream applications
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