exogenous human tgf β1  (R&D Systems)

Journal: Cells

Article Title: Human Liver Spheroids as a Model to Study Aetiology and Treatment of Hepatic Fibrosis

doi: 10.3390/cells9040964

Figure Lengend Snippet: Hepatocyte markers in monoculture and co-culture spheroids. Typical spheroid morphology was maintained across monoculture and co-culture spheroids ( a ) over 14 days. Protein expression of albumin and CYP3A4 in monoculture and co-culture spheroids from different donors was evident at day 7 ( b ). Monoculture and co-culture spheroids expressed comparable albumin and CYP3A4 protein at day 14 ( c ). mRNA expression of albumin and CYP3A4, using the same donor (n = 3), at day 7 and 14 ( d ). ALB: albumin, 3A4: CYP3A4. * p < 0.05, *** p < 0.01.

Article Snippet: Spheroids were exposed to 5 ng/mL exogenous transforming growth factor β1 (TGFβ1, 240-B-002; R&D systems, MN, USA) for 72 h on day 7, and the gene expression for TGFβ1, lysyl oxidase (LOX), and type I collagen (COL1A1) was determined.

Techniques: Co-Culture Assay, Expressing

Journal: Cells

Article Title: Human Liver Spheroids as a Model to Study Aetiology and Treatment of Hepatic Fibrosis

doi: 10.3390/cells9040964

Figure Lengend Snippet: Spheroid co-cultures express markers of NPC populations. Co-cultures stained with vimentin show dispersed localization of HSCs in different spheroid co-cultures at day 7 ( a ). The mRNA expression of vimentin was very high in co-cultures containing NPCs from donor 1 ( b ). Incorporation of vimentin-expressing cells originated from the NPC fraction with the increase in vimentin mRNA expression from the same donor combination (n = 6) being highly reproducible over the time course ( c ). Incorporation of NPCs from donor 1 was independent of the PHH donor ( d ). The presence of HSCs was further validated with the use of PDGFRβ staining ( e ) and co-staining of vimentin and αSMA on day 14, suggesting an activated HSC phenotype exclusively observed in co-cultures ( e ). VIM: Vimentin, αSMA: alpha-smooth muscle actin. PDGFRβ: platelet-derived growth factor receptor β. *** p < 0.001.

Article Snippet: Spheroids were exposed to 5 ng/mL exogenous transforming growth factor β1 (TGFβ1, 240-B-002; R&D systems, MN, USA) for 72 h on day 7, and the gene expression for TGFβ1, lysyl oxidase (LOX), and type I collagen (COL1A1) was determined.

Techniques: Staining, Expressing, Derivative Assay

Journal: Cells

Article Title: Human Liver Spheroids as a Model to Study Aetiology and Treatment of Hepatic Fibrosis

doi: 10.3390/cells9040964

Figure Lengend Snippet: Reliance on TGFβ1 expression in NPCs to produce fibrogenic outcomes. NPCs from donor 1, as compared to donor 3 NPCs, when both combined with PHH1, had higher endogenous mRNA expression of TGFβ1 ( a ) which resulted in spontaneous protein expression of αSMA and COL1A1 ( b ). Additionally, TGFβ1 protein expression was typically higher in co-cultures (donor-dependent) and could be increased under FFA pressure ( c ). Exogenous TGFβ1 in co-culture spheroids from donor 2 induced COL1A1 and LOX mRNA expression ( d ) while in donor 4, similar induction was observed under TGFβ1 pressure, which was negated by TGFβRi added before or during cultivation ( e ). ** p < 0.01.

Article Snippet: Spheroids were exposed to 5 ng/mL exogenous transforming growth factor β1 (TGFβ1, 240-B-002; R&D systems, MN, USA) for 72 h on day 7, and the gene expression for TGFβ1, lysyl oxidase (LOX), and type I collagen (COL1A1) was determined.

Techniques: Expressing, Co-Culture Assay

Journal: Cells

Article Title: Human Liver Spheroids as a Model to Study Aetiology and Treatment of Hepatic Fibrosis

doi: 10.3390/cells9040964

Figure Lengend Snippet: Regulation of fibrogenic outcomes by a TGFβ1 receptor inhibitor. Endogenous protein expression of TGFβ1 was high in donor 1 co-cultures but reduced following 72 h exposure to a TGFβRi ( a , b ) with a similar impact evident on mRNA expression of TGFβ and COL1A1 in co-cultures from donor 1 ( c ). The continuous exposure of TGFβRi for 7 days inhibited COL1A1 protein expression in a dose-dependent manner ( d ). * p < 0.05, ** p < 0.01.

Article Snippet: Spheroids were exposed to 5 ng/mL exogenous transforming growth factor β1 (TGFβ1, 240-B-002; R&D systems, MN, USA) for 72 h on day 7, and the gene expression for TGFβ1, lysyl oxidase (LOX), and type I collagen (COL1A1) was determined.

Techniques: Expressing

Journal: Cells

Article Title: Human Liver Spheroids as a Model to Study Aetiology and Treatment of Hepatic Fibrosis

doi: 10.3390/cells9040964

Figure Lengend Snippet: Effects of anti-NASH drugs on the attenuation of COL1A1 deposition in co-culture spheroids. Co-culture spheroids from donor 1 were reproducible in the fibrogenic phenotype ( a ). These spheroids were repeatedly exposed to different anti-NASH drugs, from day 7 to 14, with renewed drug exposure every alternate day. Expression of COL1A1 protein was reduced for cenicriviroc, elafibranor, and lanifibranor ( b ). Quantification (n = 10+) of αSMA and COL1A1 for each drug across replicate experiments ( c ). * p < 0.05 *, *** p < 0.001, **** p < 0.0001.

Article Snippet: Spheroids were exposed to 5 ng/mL exogenous transforming growth factor β1 (TGFβ1, 240-B-002; R&D systems, MN, USA) for 72 h on day 7, and the gene expression for TGFβ1, lysyl oxidase (LOX), and type I collagen (COL1A1) was determined.

Techniques: Co-Culture Assay, Expressing

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Endogenous versus Exogenous Growth Factor Regulation of Articular Chondrocytes

doi: 10.1002/jor.22444

Figure Lengend Snippet: Change in DNA content of articular chondrocytes in response to the designated treatments. Chondrocytes were transfected with control vectors containing 2 ug (1x) or 4 ug (2x) empty vector DNA (pAAV-MCS 1x or pAAV-MCS 2x), or vectors encoding IGF-I (pAAV-IGF-I), TGF-β1 (pAAV- TGF-β1), or both IGF-I and TGF-β1 (pAAV-IGF-I + pAAV- TGF-β1). Additional chondrocytes were transfected with control vectors and also exposed to recombinant IGF-I (pAAV-MCS 1x + rIGF-I), recombinant TGF-β1 (pAAV-MCS 1x + rTGF-β1), or both recombinant growth factors (pAAV-MCS 2x + rIGF-I + rTGF-β1). pAAV-MCS 1x was used as control for single transgenes. pAAV-MCS 2x was used as control for two transgenes. Cultures were terminated at 6 days. Data are expressed as DNA normalized to the respective control ± SD for 3 independent experiments.

Article Snippet: The exogenous TGF- β1 (R&D Systems) was recombinant mature human TGF- β1 (112 amino acids) produced in CHO cells.

Techniques: Transfection, Control, Plasmid Preparation, Recombinant

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Endogenous versus Exogenous Growth Factor Regulation of Articular Chondrocytes

doi: 10.1002/jor.22444

Figure Lengend Snippet: IGF-I produced by articular chondrocytes following the designated treatments. pAAV-MCS(1x): Empty control vector (single dose). pAAV-MCS (2x): empty control vector (double dose). pAAV-IGF-I: vector encoding IGF-I (endogenous IGF-I). pAAV-TGF-β1: vector encoding TGF-β1 (endogenous TGF-β1). rIGF-I: recombinant insulin-like growth factor I (exogenous IGF-I). rTGF-β1: recombinant transforming growth factor β1 (exogenous rTGF-β1).

Article Snippet: The exogenous TGF- β1 (R&D Systems) was recombinant mature human TGF- β1 (112 amino acids) produced in CHO cells.

Techniques: Produced, Control, Plasmid Preparation, Recombinant

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Endogenous versus Exogenous Growth Factor Regulation of Articular Chondrocytes

doi: 10.1002/jor.22444

Figure Lengend Snippet: TGF-β1 produced by articular chondrocytes following the designated treatments. pAAV-MCS(1x): Empty control vector (single dose). pAAV-MCS (2x): empty control vector (double dose). pAAV-IGF-I: vector encoding IGF-I (endogenous IGF-I). pAAV-TGF-β1: vector encoding TGF-β1 (endogenous TGF-β1). rIGF-I: recombinant insulin-like growth factor I (exogenous IGF-I). rTGF-β1: recombinant transforming growth factor β1 (exogenous rTGF-β1).

Article Snippet: The exogenous TGF- β1 (R&D Systems) was recombinant mature human TGF- β1 (112 amino acids) produced in CHO cells.

Techniques: Produced, Control, Plasmid Preparation, Recombinant