recombinant fgf4  (R&D Systems)

Journal: Genes & Development

Article Title: Transcriptional derepression of negative regulators of MAP kinase supports maintenance of diapause ES cells in the pluripotent state

doi: 10.1101/gad.353143.125

Figure Lengend Snippet: The differentiation block in I-BET-resistant (I-BETR) diapause-like ES cells. ( A ) Alkaline phosphatase (AP) levels in control and I-BETR ES cells. Scale bar, 100 µm. ( B ) Expression levels of the pluripotency ( left panel) or differentiation-inducing ( right panel) genes in control, 2i-treated, or I-BETR ES cells. RNA expression levels were quantified by qPCR. Values represent normalized mean ± SD. n = 3. ( C ) Colony morphology and/or alkaline phosphatase (AP) levels in control and I-BETR ES cells following vehicle or FGF4 triggering. Scale bar, 100 µm. The bar graph represents quantification of relative percentage of pluripotent colonies in different groups. Values represent ± SD. n = 9. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( D ) Expression levels of selected pluripotency ( left ) or differentiation-inducing ( right ) genes in control and I-BETR ES cells treated or not treated with FGF4. Error bars indicate SD. n = 3. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( E ) Withdrawal of I-BET (I-BETW) restores the control ES cell-like pluripotency gene expression pattern in I-BETR ES cells. Values represent gene expression levels normalized to the mean of control samples based on TPM values observed by bulk mRNA RNA-seq analysis . ( F ) Generation of chimeras by I-BETR ES cells. The I-BETR ES cells were incubated in I-BET-free medium for 12–14 h and injected into the C57BL/6J blastocysts. White coat color indicates the chimerism.

Article Snippet: For FGF4-driven ES cell differentiation, 10 ng/μL recombinant FGF4 (R&D Systems 235-F4) was added on day 0 together with 1 μg/μL heparin (Sigma-Aldrich H3149).

Techniques: Blocking Assay, Control, Expressing, RNA Expression, Gene Expression, RNA Sequencing, Incubation, Injection

Journal: Biology of the Cell

Article Title: Generation of Intestinal and Colonic Organoids Derived From Human Pluripotent Stem Cells

doi: 10.1111/boc.70044

Figure Lengend Snippet: Overview of the protocol steps. The complete directed differentiation process lasts for 37 days, including 28 days of culture in Matrigel. Two days after seeding the hPSCs, the first step involves adding Activin A for 3 days to generate definitive endoderm formation, followed by the addition of FGF4 and CHIRON for 4 more days to induce spheroid formation. The resulting spheroids are then embedded in Matrigel and cultured for 28 days, with a passage at day 14 to renew the Matrigel.

Article Snippet: FGF4 (Human recombinant fibroblast growth factor 4 premium grade, Miltenyi Biotec, cat. no. 130‐109‐391) ▲CRITICAL .

Techniques: Cell Culture

Journal: Biology of the Cell

Article Title: Generation of Intestinal and Colonic Organoids Derived From Human Pluripotent Stem Cells

doi: 10.1111/boc.70044

Figure Lengend Snippet: Characterization of definitive endoderm induction and spheroids generation. (a) Immunofluorescence images for FOXA2 and SOX17 of definitive endoderm after 3 days of Activin A induction. (b) Immunofluorescence images of hindgut spheroids following 4 days of FGF4 and Chiron induction, characterized by CDX2 (intestinal marker) and E‐cadherin (epithelial marker) expression. Scale bars = 100 µm.

Article Snippet: FGF4 (Human recombinant fibroblast growth factor 4 premium grade, Miltenyi Biotec, cat. no. 130‐109‐391) ▲CRITICAL .

Techniques: Immunofluorescence, Marker, Expressing