Journal: The International journal of developmental biology

Article Title: Expression of NANOG and NANOGP8 in a variety of undifferentiated and differentiated human cells

doi: 10.1387/ijdb.103192sa

Figure Lengend Snippet: HUMAN CELL TYPES/RNA USED IN NANOG TRANSCRIPTION ANALYSIS

Article Snippet: Identification of NANOG transcripts in both differentiated and undifferentiated human cells Based upon our previously published work in which we detected the expression of NANOG in a primary adult human fibroblast culture ( Page et al. , 2009 ) we sought to examine NANOG expression across a panel of differentiated and undifferentiated cell types: human adult fibroblasts, human neonatal fibroblasts, HeLa cells, SH-SY5Y neuroblastoma cells, human smooth muscle cells (hSMCs), human vascular endothelial cells (HUVECs), mesenchymal stem cells (hMSCs), total heart tissue, and H9 human embryonic stem cells (hESCs) ( ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Cell type Source Catalog number Adult fibroblasts ATCC CRL-2352 Embryonic fibroblasts ATCC CRL-2097 HeLa cells ATCC CCL-2 SH-SY5Y Neuroblastoma cells ATCC CRL-2266 Smooth muscle cells (hSMCs) Dr. B. Blackman, U. Virginia NA Umbilical vein endothelial cells (HUVECs) Lonza CC-2517 Mesenchymal stem cells (hMSCs) Lonza AA-2501 Total Heart (RNA) ClonTech 636532 H9 Embryonic stem cells (hESCs) WiCell WA09 Human teratocarcinoma cells ATCC CRL-2073 Open in a separate window HUMAN CELL TYPES/RNA USED IN NANOG TRANSCRIPTION ANALYSIS We analyzed the presence of NANOG transcript by quantitative PCR (qPCR, primers listed in ).

Techniques:

Journal: bioRxiv

Article Title: Rostrocaudal patterning and neural crest differentiation of human pre-neural spinal cord progenitors in vitro

doi: 10.1101/2020.06.16.155564

Figure Lengend Snippet: A, B) Optimal CHIR concentration (3 μM) was optimised in WA01 (H1) hESCs. (A) Representative immunostaining of NMP markers SOX2 (red) and CDX2 (grey) and TBXT (magenta) at 36h after following treatment scheme with 3 μM CHIR and (B) quantification markers over a range of CHIR concentrations between 1-10 μM. Scale bars, 100μm. C,D) Transcriptional analysis (RT-qPCR) of NMP markers TBXT, SOX2 and CDX2 (C) and selected HOX genes (D) up to passage 10. Expression levels are normalised to the reference gene PBDG . Error bars show SD, (n=3 technical replicates). E, F) Optimal CHIR concentration (4 μM) was optimised in AICS ZO1-mEGFP (AICS-0024) iPSCs hESCs. (E) Representative immunostaining of NMP markers SOX2 (red) and CDX2 (grey) and TBXT (magenta) at 36h after following treatment scheme with 3 μM CHIR and (F) quantification markers over a range of CHIR concentrations between 1-10 μM. Scale bars, 100μm. G,H) Transcriptional analysis (RT-qPCR) of NMP markers TBXT, SOX2 and CDX2 (G) and selected HOX genes (H) up to passage 10. Expression levels are normalised to the reference gene PBDG . Error bars show SD, (n=3 technical replicates).

Article Snippet: Human ESCs (WA09 and WA01, WiCell) and human iPSCs (AICS-23, Allen Institute) were maintained in feeder-free cultures, plated on Corning Matrigel Growth Factor Reduced (GFR) Basement Membrane Matrix (Corning Incorporated, 354230) and grown in mTESR1 (STEMCELL technologies, 85850).

Techniques: Concentration Assay, Immunostaining, Quantitative RT-PCR, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Sex-biased gene expression during neural differentiation of human embryonic stem cells

doi: 10.3389/fcell.2024.1341373

Figure Lengend Snippet: Similarities between male and female hESC in neuronal differentiation: (A) Overview of the experimental procedure: Four hESC lines of each sex were differentiated for 37 days to a population of mixed neurons induced by small molecules in 2D culture. Bulk RNA sequencing and qPCR. (B) A PCA analysis of expressed genes displays similar gene expression patterns at D0, some inter-cell line differences during early differentiation, and a sex-biased separation at D37 (XY: WA14, HS401, HS983a, XX: WA09, HS975, HS980). (C) Marker gene expression for pluripotency (OCT4, NANOG), ectoderm (PAX6, SOX1), immature neurons (DCX, NCAM), mature neurons (MAP2, DLG4, SYP), post-mitotic neurons (ELAVL2, TUBA1A, NDN), as well as for proliferation (MKI67, PCNA), and apoptosis (CASP3, BCL2) show a similar expression in male and female cell lines. (D) Markers for specialized neurons (GABAergic, Dopaminergic, Glutamatergic) display a premature state of the neurons after 37 days of differentiation. (E) A gene set enrichment analysis at D4, D9, and D37 reveals an upregulation of biological processes implicated in neuronal differentiation in all cell lines. The GSEA was summarized in an enrichment map. Each dot represents a gene set; the size of the dot corresponds to the size of the gene set. If the dots are connected or in close proximity, the more the gene sets are overlapping, and the closer related are their associated biological functions. Gene sets were grouped by a common GO-term classification and named after an overarching biological function. Biological functions implicated in neuronal development are highlighted in blue. Gene sets are filtered by FDR q-values <0.1. created with BioRender.com .

Article Snippet: The human embryonic stem cell lines WA14 p33 (XY) ( ) and WA09 LT2e-H9CAGGFP p80 (XX) ( ) were acquired from WiCell, Madison, Wisconsin.

Techniques: RNA Sequencing Assay, Expressing, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: Sex-biased gene expression during neural differentiation of human embryonic stem cells

doi: 10.3389/fcell.2024.1341373

Figure Lengend Snippet: hESC lines and gene expression analysis method and timepoints.

Article Snippet: The human embryonic stem cell lines WA14 p33 (XY) ( ) and WA09 LT2e-H9CAGGFP p80 (XX) ( ) were acquired from WiCell, Madison, Wisconsin.

Techniques: Expressing