Journal: Nature Communications

Article Title: Expansion of functional personalized cells with specific transgene combinations

doi: 10.1038/s41467-018-03408-4

Figure Lengend Snippet: Overview of cell types immortalized in this study

Article Snippet: The following media were used for cultivation of fibroblasts: huFib medium (InSCREENeX GmbH), bone marrow stroma cells: huMSC medium (InSCREENeX GmbH), HUVECs: huVEC medium (InSCREENeX GmbH), HMVECs: huMEC medium (InSCREENeX GmbH), LECs: huLEC medium (InSCREENeX GmbH), chondrocytes: huChon medium (InSCREENeX GmbH), and osteoblasts: huOB maintenance medium (InSCREENeX GmbH).

Techniques:

Journal: Nature Communications

Article Title: Expansion of functional personalized cells with specific transgene combinations

doi: 10.1038/s41467-018-03408-4

Figure Lengend Snippet: Hepatocyte cell lines retain key hepatic properties. a Albumin and Glucose-6-phosphatase gene expression analysis of expandable hepatocytes cultured in standard 2D or 3D (9 days of cultivation as spheroids in spinner flasks) conditions and primary hepatocytes (pHep) either cultured in 3D conditions or freshly isolated (expression normalized to Gapdh, n = 3 independent experiments, except for pHep were n = 2 independent experiments, bars represent mean). b Immunofluorescence-based detection of Albumin and E-cadherin in e-mHepA expandable hepatocytes counterstained for nuclei with DAPI. Scale bar, 20 μm. c Glycogen storage in expandable hepatocytes and control fibroblasts as analyzed by Periodic Acid-Schiff staining. Representative bright field micrographs and quantification of multiple field of views are shown. Scale bar, 100 μm (mean, pooled data from n = 3 independent experiments, field of views analyzed in total 35 to 42). d Immunofluorescence micrograph of Albumin and Hnf4α in e-mHepB hepatocytes after 3D culture as spheroids for 9 days. Actin filaments and nuclei were counterstained with phalloidin and DAPI, respectively. Scale bar, 20 µm. e Induction of phase I metabolic enzyme expression in expandable hepatocytes cultured in 2D or 3D conditions and control fibroblasts in response to stimulation with 50 μM Dexamethasone or 2 μM 3-Methylcholanthrene for 72 h (expression normalized to Gapdh, n = 3 independent experiments, except for e-mHepB Cyp1a1 expression were n = 6 independent experiments, bars represent mean). f Luminescence-based detection of Phase I enzymatic activity in expandable hepatocytes and control fibroblasts after cells were treated as indicate above (activity normalized to 10 4 cells, pooled data from n = 2 independent experiments with n = 3 technical replicates each). g Staining of livers of Fah –/– / Rag2 –/– /Il2rg –/– (FRG) mice for FAH and GFP, 90 days after transplantation of GFP-tagged expandable hepatocytes. Scale bar, 100 µm

Article Snippet: The following media were used for cultivation of fibroblasts: huFib medium (InSCREENeX GmbH), bone marrow stroma cells: huMSC medium (InSCREENeX GmbH), HUVECs: huVEC medium (InSCREENeX GmbH), HMVECs: huMEC medium (InSCREENeX GmbH), LECs: huLEC medium (InSCREENeX GmbH), chondrocytes: huChon medium (InSCREENeX GmbH), and osteoblasts: huOB maintenance medium (InSCREENeX GmbH).

Techniques: Gene Expression, Cell Culture, Isolation, Expressing, Immunofluorescence, Control, Staining, Activity Assay, Transplantation Assay

Journal: Nature Communications

Article Title: Expansion of functional personalized cells with specific transgene combinations

doi: 10.1038/s41467-018-03408-4

Figure Lengend Snippet: Comparable phenotype of primary and expanded endothelial cells. a Phase contrast microscopy shows the expandable HUVEC cell line e-hUVEC-2 (MYC, ID1, and ID2). Scale bar 100 µm. b CD31 expression of human endothelial cell populations immortalized with three different gene sets as indicated. c Global gene expression analysis was performed on three different HUVEC lines (e-hUVEC-2, 8 and 9; in duplicate) (3, 4, 5, respectively) and two independent primary HUVEC populations (2) as well as four primary gingiva fibroblast populations (1). Expression data was processed with GeneSpring 11.5.1 software and a Standard Pearson Correlation was determined for each gene versus all other genes. The correlation heatmap depicts the pair-wise correlation coefficient between the given samples and displays the relationship between the different samples. The samples are clustered based on the pair-wise correlation coefficients between all entities. d Phenotypic stability of e-hUVEC-2 cells after 45 and 90 cumulative population doublings was evaluated by CD31 (also known as PECAM1) expression, acetylated LDL uptake, and eNOS activity. Gray fill: antibody isotype control; black outline: stained sample. MFI: median fluorescence intensity. e Immunofluorescence-based detection of CD31 and CD146 (also known as MCAM) in expandable HUVECs counterstained for nuclei with DAPI. Scale bars, 100 µm. f The phenotype and the functionality of cell line e-hUVEC-2 (cumulative population doubling 80) was compared to primary HUVECs based on flow cytometric analysis of CD31, TIE1, TIE2, and CD309 (also known as VEGFR2) expression. Gray fill: antibody isotype control; black outline: stained sample. MFI: median fluorescence intensity. g The angiogenic potential of primary HUVECs and e-hUVEC-2 was determined in vitro by a matrigel tube formation assay. Scale bars, 200 µm. h Spheroids in matrigel of primary HUVEC and e-hUVEC-2 were subcutaneously injected into Rag2 -/- Il2r g −/− mice. After two weeks the implants were dissected and stained for human CD31 (brown color). e-hUVEC-2 organized into human CD31 positive microvessels similar to primary HUVEC. Scale bars, 100 µm

Article Snippet: The following media were used for cultivation of fibroblasts: huFib medium (InSCREENeX GmbH), bone marrow stroma cells: huMSC medium (InSCREENeX GmbH), HUVECs: huVEC medium (InSCREENeX GmbH), HMVECs: huMEC medium (InSCREENeX GmbH), LECs: huLEC medium (InSCREENeX GmbH), chondrocytes: huChon medium (InSCREENeX GmbH), and osteoblasts: huOB maintenance medium (InSCREENeX GmbH).

Techniques: Microscopy, Expressing, Gene Expression, Software, Activity Assay, Control, Staining, Fluorescence, Immunofluorescence, In Vitro, Tube Formation Assay, Injection

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: cGMP accumulation in co‐cultures of human primary vascular smooth muscle cells following addition of serelaxin to endothelium. HUAEC , HUVEC or HCAEC were co‐cultured with (A) HUASMC or (B) HUVSMC (all n = 5), and the ECs were treated with serelaxin for 30 min. Serelaxin addition to HUAEC did not cause cGMP accumulation in HUAEC (▲) (C) HUASMC (□) or (D) HUVSMC (◯) co‐cultured with HUAEC, whereas direct stimulation of either (C) HUASMC (n = 5) or (D) HUVSMC with serelaxin caused a concentration‐dependent increase in cGMP accumulation (dashed lines). In contrast, serelaxin addition to HUVEC concentration‐dependently increased cGMP accumulation not only in HUVEC (■) but also in (E) HUASMC (□) or (F) HUVSMC (◯) co‐cultured with HUVEC with the responses in smooth muscle cells being greater or in the case of HUVSMC much greater than cGMP responses to direct stimulation of (E) HUASMC or (F) HUVSMC (dashed lines). A similar pattern of cGMP accumulation was observed with (G, H) HCAEC (●) and (G) HUASMC (□) or (H) HUVSMC (◯) co‐cultured with HCAEC.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Concentration Assay

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: cAMP accumulation in co‐cultures of human primary vascular smooth muscle cells following addition of serelaxin to endothelium (all n = 5). HUAEC, HUVEC or HCAEC were co‐cultured with (A) HUASMC or (B) HUVSMC, and the endothelial cells were treated with serelaxin for 30 min. Serelaxin added to HUAEC did not cause cAMP accumulation either in (C, D) HUAEC (▲), (C) HUASMC (□) or (D) HUVSMC (◯), whereas direct stimulation of (C) HUASMC or (D) HUVSMC with serelaxin caused a concentration‐dependent increase in cAMP accumulation (dashed lines). Although direct addition of serelaxin to HUVEC concentration‐dependently increased cAMP accumulation in (E, F) HUVEC (■), there was no significant effect on cAMP accumulation in (E) HUASMC (□) or (F) HUVSMC (◯). Direct addition of serelaxin to (E) HUASMC or (F) HUVSMC stimulated cAMP accumulation (dashed lines). Serelaxin concentration‐dependently increased cAMP accumulation in (G, H) HCAEC (●) but also caused a robust concentration‐dependent increase in cAMP accumulation in both (G) HUASMC (□) and (H) HUVSMC (◯).

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Concentration Assay

Journal: Toxicology Research

Article Title: Nanoparticle induced barrier function assessment at liquid–liquid and air–liquid interface in novel human lung epithelia cell lines † †Electronic supplementary information (ESI) available. See DOI: 10.1039/c9tx00179d

doi: 10.1039/c9tx00179d

Figure Lengend Snippet: Characterization of growth behavior: Growth curves of A549, hAELVi and huAEC cells show similar growth behavior for all cell types.

Article Snippet: CI-hAELVi (cat. no.: INS-CI-1015) and CI-huAEC (cat. no.: INS-CI-1011) cells were purchased from InSCREENeX GmbH (InSCREENeX GmbH, Germany).

Techniques: