Journal: Frontiers in Genetics

Article Title: A Human Gonadal Cell Model From Induced Pluripotent Stem Cells

doi: 10.3389/fgene.2018.00498

Figure Lengend Snippet: List of genes from expressed in the different stages of differentiation and maturation of SCs based on literature search.

Article Snippet: Human foreskin fibroblast (HFFn, PC501 A-HFF, System Biosciences Mountain View, CA, United States) were cultured in DMEM medium supplemented with 10% FBS and 1% Pen/Strep according to the manufacturer instructions.

Techniques: Wilms Tumor Assay, Binding Assay

Journal: Frontiers in Genetics

Article Title: A Human Gonadal Cell Model From Induced Pluripotent Stem Cells

doi: 10.3389/fgene.2018.00498

Figure Lengend Snippet: Graphic representation of Human Fibroblasts (HFs) cell reprogramming and iPSCs differentiation toward SLCs. AP, alkaline phosphatase; EBs, embrioid bodies; FGF9, fibroblast growth factor 9; IF, immunofluorescence; iPSCs, induced pluripotent stem cells; PGD2, prostaglandin 2; RNA-seq, next generation RNA sequencing; SLCs, Sertoli-like cells; SNL, SNL feeder cells.

Article Snippet: Human foreskin fibroblast (HFFn, PC501 A-HFF, System Biosciences Mountain View, CA, United States) were cultured in DMEM medium supplemented with 10% FBS and 1% Pen/Strep according to the manufacturer instructions.

Techniques: Immunofluorescence, RNA Sequencing

Journal: Frontiers in Genetics

Article Title: A Human Gonadal Cell Model From Induced Pluripotent Stem Cells

doi: 10.3389/fgene.2018.00498

Figure Lengend Snippet: (A) Immunofluorescence staining of iPSC colonies. Zoom-in details in right-down squares. Cells were stained with antibodies against OCT4, TRA-1-81, SSEA3, SSEA4, or SSEA1 and SOX2. Mounting medium contains DAPI stain. Pluripotency markers OCT4, TRA-1-81, and SOX2 positively marked the nuclei of cells. SSEA3 and SSEA4 were localized in the cytoplasmic area of colonies. SSEA1 expression was absent in colony nuclei but observed in the feeder cells. (B) RT-PCR analysis of pluripotential colonies for the genes DPPA2, DPPA4, ESG1, SOX2, and NANOG. Primers used for SOX2 and NANOG specifically detect transcripts from the endogenous genes, but not from the lentiviral transgenes. Four (1–4) different colonies are shown. HF, terminally differentiated fibroblasts; iPSC, induced pluripotent stem cells. (C,D) Teratoma formation analysis. Histology sections of teratoma stained with Hematoxylin & eosin. 1: mesoderm, 2: endoderm, 3: ectoderm. Scale bars 50 μm for (A,C) , and 100 μm for (D) .

Article Snippet: Human foreskin fibroblast (HFFn, PC501 A-HFF, System Biosciences Mountain View, CA, United States) were cultured in DMEM medium supplemented with 10% FBS and 1% Pen/Strep according to the manufacturer instructions.

Techniques: Immunofluorescence, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Genetics

Article Title: A Human Gonadal Cell Model From Induced Pluripotent Stem Cells

doi: 10.3389/fgene.2018.00498

Figure Lengend Snippet: Immunofluorescence staining of SLC colonies with antibodies against SOX9. Mounting medium contains DAPI stain. SOX9 were expressed in SLC colonies. HF, terminally differentiated fibroblasts; HSerCs, human Sertoli cells; NT2, NT2d1 cells; SOX9, SRY (sex determining region Y)-box 9. Scale bars 50 μm.

Article Snippet: Human foreskin fibroblast (HFFn, PC501 A-HFF, System Biosciences Mountain View, CA, United States) were cultured in DMEM medium supplemented with 10% FBS and 1% Pen/Strep according to the manufacturer instructions.

Techniques: Immunofluorescence, Staining

Journal: Frontiers in Genetics

Article Title: A Human Gonadal Cell Model From Induced Pluripotent Stem Cells

doi: 10.3389/fgene.2018.00498

Figure Lengend Snippet: Secreted AMH quantification by ELISA. SLCs secreted AMH levels were significantly higher when compared with AMH secretion by HSerCs or NT2d1 cells. HF: terminally differentiated fibroblasts, HSerCs: primary human Sertoli cells, NT2d1: NT2d1 cells, SLCs: Sertoli like cells. n = 4 for SLCs and HSerCs, n = 2 for HFs and NT2d1. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗∗ P < 0.0001.

Article Snippet: Human foreskin fibroblast (HFFn, PC501 A-HFF, System Biosciences Mountain View, CA, United States) were cultured in DMEM medium supplemented with 10% FBS and 1% Pen/Strep according to the manufacturer instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Cell Cycle

Article Title: Post-irradiation promotes susceptibility to reprogramming to pluripotent state in human fibroblasts

doi: 10.1080/15384101.2017.1371887

Figure Lengend Snippet: Characterization of induced pluripotent stem cells (iPSCs) generated from post-irradiated fibroblasts using four factors (OSKM) or three factors (OSK). (A) Morphology, immunostaining for pluripotent markers (Oct4, Sox2, and Nanog), and alkaline phosphatase (ALP) assay in 4F- or 3F-iPSCs (passage 7) derived from 5Gy-D14 cells. DAPI was used for counting nuclear staining. Scale bar = 100 μm. (B) Quantitative RT-PCR analysis of expression of endogenous pluripotent markers (Nanog, Oct4, and Sox2) in 4F- or 3F-iPSCs (5Gy-D14). mRNA expression levels of pluripotent markers was calculated relative to that of GAPDH and normalized to the parental cells as the control. The data are shown as means ± SDs from triplicate experiments (**P < 0.01, one-way ANOVA analysis with Scheffe pairwise post-hoc test) (C) DNA methylation analysis of several CpG sites in Oct4 and Nanog promoters, indicating demethylation of Oct4 and Nanog promoters in 4F- or 3F-iPSCs generated from 5Gy-D14 cells. Oct4 and Nanog promoters of parental cells were hyper-methylated. (D) Karyotyping of 4F- or 3F-iPSCs generated from 5Gy-D14 cells. NUFF: newborn human foreskin fibroblasts as donor cells. 4F-iPSC (5Gy-D14): iPSCs derived from fibroblasts at 14 d post-irradiation with 5 Gy using four factors (OSKM). 3F-iPSC (5Gy-D14): iPSCs derived from fibroblasts at 14 d post-irradiation with 5 Gy using three factors (OSK).

Article Snippet: Newborn human foreskin fibroblasts (passage 9) were purchased from Globalstem Inc. and maintained in DMEM containing 10% FBS.

Techniques: Generated, Irradiation, Immunostaining, ALP Assay, Derivative Assay, Staining, Quantitative RT-PCR, Expressing, DNA Methylation Assay, Methylation