embryonic stem cell esc line h9  (WiCell Research Institute Inc)

Journal: Heliyon

Article Title: REST and RCOR genes display distinct expression profiles in neurons and astrocytes using 2D and 3D human pluripotent stem cell models

doi: 10.1016/j.heliyon.2024.e32680

Figure Lengend Snippet: REST and RCOR gene expression in functional and mature NGN2 iNs. (A) hPSCs were differentiated into functionally mature iNs derived from H9 ESCs and two healthy control iPSC lines (iPSC1 and iPSC2) using NGN2 overexpression. (B ) Immunocytochemistry confirmed expression of MAP2 and VGLUT1 at a protein level in neurons derived from all cell lines (n = 3 independent differentiations). Scale bars = 50 μm. (C) Representative current clamp recordings of membrane potential responses of the H9, iPSC1 and iPSC2 cell lines (n = 16, 12 and 16 cells, respectively, from 3 independent differentiations). The stimulation protocols consisted of 1 s long current injections from −100 pA to 100 pA in 10 pA steps. The coloured traces indicate firing activity at rheobase. (D) REST and RCOR gene relative expression levels were analysed via RT-qPCR. Data was calculated as the mean of three housekeeping genes and presented as mean ± SEM. Data was analysed using an ordinary Two-way ANOVA with a Holm-Sidak test to correct for multiple comparisons. * p < 0.05, *** p < 0.001. n = 3 biological replicates, with each data point representing the average of 3 technical replicates.

Article Snippet: The H9 human embryonic stem cells (ESCs) line (WA09 obtained from WiCell; referred to as ESC) and two healthy control induced pluripotent stem cell (iPSC) lines, iPSC1 [ ] and iPSC2 [ ] were utilised in the current study. hPSCs were cultured as previously reported in Abu-Bonsrah et al. [ ], Denham and Dottori [ ] and Mattei et al. [ ].

Techniques: Expressing, Functional Assay, Derivative Assay, Control, Over Expression, Immunocytochemistry, Membrane, Activity Assay, Quantitative RT-PCR