Balancing serendipity and reproducibility: Pluripotent stem cells as experimental systems for intellectual and developmental disorders

Abstract

Reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) and their differentiation into neural lineages is a revolutionary experimental system for studying neurological disorders, including intellectual and developmental disabilities (IDDs). However, issues related to variability and reproducibility have hindered translating preclinical findings into drug discovery. Here, we identify areas for improvement by conducting a comprehensive review of 58 research articles that utilized iPSC-derived neural cells to investigate genetically defined IDDs. Based upon these findings, we propose recommendations for best practices that can be adopted by research scientists as well as journal editors.

Generation of a NES-mScarlet Red Fluorescent Reporter Human iPSC Line for Live Cell Imaging and Flow Cytometric Analysis and Sorting Using CRISPR-Cas9-Mediated Gene Editing

“…Normalized ratios for each gene of interest were calculated by the formula 2 −dCt , and normalized fold changes (relative to undifferentiated hiPSCs) were calculated by the 2 −ddCt method, in which dCt is the Ct value of the gene of interest minus the Ct value of the housekeeping gene ACTB (internal control) [ 27 ]. Data are from three technical replicates (independent differentiation experiments) [ 28 ].…”

“…Data are represented as the mean ± SEM from three or two independent experiments (technical replicates, [ 28 ]) for RT-qPCR and cell counting or ELISA, respectively. For statistical calculations, graphs were plotted and a two-sample t -test was conducted with a significance threshold of 95% (* p < 0.05) or 99.5% (** p < 0.005) using MATLAB software version R2021b.…”

What Molecular Recognition Systems Do Mesenchymal Stem Cells/Medicinal Signaling Cells (MSC) Use to Facilitate Cell-Cell and Cell Matrix Interactions? A Review of Evidence and Options

“…Recently. Such iPSC have been investigated for the potential treatment of heart diseases (discussed in [85]), neurodegenerative conditions (discussed in [86][87][88][89]), and musculoskeletal conditions such as intervertebral disc repair (reviewed in [90]).…”

Tips and tricks for successfully culturing and adapting human induced pluripotent stem cells

“… 21 , 22 It is worth mentioning that the problems that we had to face during the process of generation and characterization of the cells were numerous, especially regarding the adaptation to feeder-free conditions. There are currently a lot of published combinations of matrices and culture media to replace feeder layers (reviewed in Dakhore et al., 8 Sams and Powers, 8 , 11 , 13 , 20 , 23 , 24 , 25 Skottman and Hovatta, 8 , 11 , 13 , 20 , 23 , 24 , 25 Liu et al., 8 , 11 , 13 , 20 , 23 , 24 , 25 Anderson et al., 8 , 11 , 13 , 20 , 23 , 24 , 25 Nakamura et al., 8 , 11 , 13 , 20 , 23 , 24 , 25 Kishimoto et al. 8 , 11 , 13 , 20 , 23 , 24 , 25 ).…”

“…There are currently a lot of published combinations of matrices and culture media to replace feeder layers (reviewed in Dakhore et al., 8 Sams and Powers, 8 , 11 , 13 , 20 , 23 , 24 , 25 Skottman and Hovatta, 8 , 11 , 13 , 20 , 23 , 24 , 25 Liu et al., 8 , 11 , 13 , 20 , 23 , 24 , 25 Anderson et al., 8 , 11 , 13 , 20 , 23 , 24 , 25 Nakamura et al., 8 , 11 , 13 , 20 , 23 , 24 , 25 Kishimoto et al. 8 , 11 , 13 , 20 , 23 , 24 , 25 ). But when consulting the bibliography for troubleshooting, we could just find depthless explained methodology.…”