Journal: bioRxiv

Article Title: 3D-bioprinted, phototunable hydrogel models for studying adventitial fibroblast activation in pulmonary arterial hypertension

doi: 10.1101/2022.11.11.516188

Figure Lengend Snippet: Fibroblast activation in 3D-bioprinted models of pulmonary arterial adventitia. A) Fibrotic activation in soft and stiffened 3D hydrogels measured by αSMA expression. HPAAFs in stiffened constructs were significantly more positive for αSMA than cells in soft constructs. Columns represent mean ± SEM, n = 3. *, p < 0.05, Mann-Whitney U test. B) Representative confocal images of immunostaining for αSMA, actin, and DAPI in soft and stiffened 3D hydrogels. HPAAFs in stiffened constructs showed more prevalent αSMA immunofluorescence than cells in soft constructs. Scale bar = 250 µm. C) Fibroblast proliferation in soft and stiffened 3D bioprinted constructs measured by EdU positivity. HPAAFs in stiffened constructs were significantly more positive for EdU than cells in soft constructs. Columns represent mean ± SEM, n = 3. *, p < 0.05, Mann-Whitney U test. D) Representative confocal images of immunostaining for EdU and Hoechst dye in soft and stiffened 3D hydrogels. HPAAFs in stiffened constructs showed more prevalent EdU immunofluorescence than cells in soft constructs. Scale bar = 300 µm.

Article Snippet: [ ] Human pulmonary artery adventitia fibroblasts (HPAAFs; Accegen, Fairfield, NJ) from a 2-year-old male patient were cultured in complete HPAAF culture media according to manufacturer instructions.

Techniques: Activation Assay, Expressing, Construct, MANN-WHITNEY, Immunostaining, Immunofluorescence

Journal: Cell Proliferation

Article Title: A novel multikinase inhibitor SKLB‐YTH‐60 ameliorates inflammation and fibrosis in bleomycin‐induced lung fibrosis mouse models

doi: 10.1111/cpr.13081

Figure Lengend Snippet: YTH‐60 inhibits fibroblast activation and proliferation. A‐C, NIH‐3T3, HPF and MRC‐5 were treated with different concentrations of YTH‐60 and Nintedanib for 24, 48 or 72 h and cell viability was measured by the CCK8 assay. The values are expressed as the mean ± SD (n = 3); * P < .05; ** P < .01; *** P < .001 compared with YTH‐60 control; # P < .05; ## P < .01; ### P < 0.001 compared with Nintedanib control. D‐F, Real‐time qPCR analysis of collagen Ⅰ, α‐SMA and TGF‐β1 in NIH‐3T3 cells treated with 5 ng·mL −1 TGF‐β1 and 2.5 μmol·L −1 YTH‐60 or Nintedanib for 24 h. The values are expressed as the mean ± SD, n = 3; * P < .05; ** P < .01; *** P < .001. G‐I, NIH‐3T3, HPF and MRC‐5 were stimulated with TGF‐β1 by 2.5 and 5 μmol·L −1 YTH‐60 administration for 24 h, and expression of α‐SMA and collagen Ⅰ using western blot. J‐K, expression of α‐SMA and collagen Ⅰ was visualized by immunofluorescence, Scale bar = 50 μm

Article Snippet: Human pulmonary fibroblasts (HPF) was purchased from Sciencell.

Techniques: Activation Assay, CCK-8 Assay, Control, Expressing, Western Blot, Immunofluorescence

Journal: Aging (Albany NY)

Article Title: c-Met as a new marker of cellular senescence

doi: 10.18632/aging.101961

Figure Lengend Snippet: The levels of c-Met and pMet proteins in primary cultures of human dermal (left panel) and pulmonary fibroblasts (right panel) of various passages. Immunoblots represent one of the 4 independent experiments.

Article Snippet: Primary cultures of human dermal and pulmonary fibroblasts (obtained from ScienCell, Carlsbad, CA, US) were grown under standard conditions (37 °C, 5% CO 2 ) in Dulbecco’s modified Eagles medium (DMEM), supplemented with 10% fetal bovine serum, 1% L-glutamine, and 1% penicillin/streptomycin.

Techniques: Western Blot

Journal: Aging (Albany NY)

Article Title: c-Met as a new marker of cellular senescence

doi: 10.18632/aging.101961

Figure Lengend Snippet: The levels of Akt1/2/3 and pAkt proteins in primary cultures of human dermal (left panel) and pulmonary fibroblasts (right panel) of various passages. Immunoblots represent one of the 3 independent experiments.

Article Snippet: Primary cultures of human dermal and pulmonary fibroblasts (obtained from ScienCell, Carlsbad, CA, US) were grown under standard conditions (37 °C, 5% CO 2 ) in Dulbecco’s modified Eagles medium (DMEM), supplemented with 10% fetal bovine serum, 1% L-glutamine, and 1% penicillin/streptomycin.

Techniques: Western Blot

Journal: Aging (Albany NY)

Article Title: c-Met as a new marker of cellular senescence

doi: 10.18632/aging.101961

Figure Lengend Snippet: The levels of Stat3 proteins in primary cultures of human dermal (left panel) and pulmonary fibroblasts (right panel) of various passages. Immunoblots represent one of the 3 independent experiments.

Article Snippet: Primary cultures of human dermal and pulmonary fibroblasts (obtained from ScienCell, Carlsbad, CA, US) were grown under standard conditions (37 °C, 5% CO 2 ) in Dulbecco’s modified Eagles medium (DMEM), supplemented with 10% fetal bovine serum, 1% L-glutamine, and 1% penicillin/streptomycin.

Techniques: Western Blot

Journal: Aging (Albany NY)

Article Title: c-Met as a new marker of cellular senescence

doi: 10.18632/aging.101961

Figure Lengend Snippet: Evidence for the involvement of Akt and Stat3 in cellular senescence.

Article Snippet: Primary cultures of human dermal and pulmonary fibroblasts (obtained from ScienCell, Carlsbad, CA, US) were grown under standard conditions (37 °C, 5% CO 2 ) in Dulbecco’s modified Eagles medium (DMEM), supplemented with 10% fetal bovine serum, 1% L-glutamine, and 1% penicillin/streptomycin.

Techniques: Activity Assay

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Silencing of Sodium-Hydrogen Exchanger 1 Attenuates the Proliferation, Hypertrophy, and Migration of Pulmonary Artery Smooth Muscle Cells via E2F1

doi: 10.1165/rcmb.2011-0032OC

Figure Lengend Snippet: Effects of silencing NHE1 on proliferation of human (HPASMCs), human pulmonary artery endothelial cells (HPAECs), and human pulmonary artery fibroblasts (HPAFs) under condition of hypoxia. (A) Representative RT-PCR data show expression of NHE1 mRNA in three cell types. RNA was isolated from HPASMCs, HPAECs, and HPAFs. RT-PCR was performed to measure the expression of NHE1. GAPDH was used as loading control. (B) Cell proliferation data. After transfection with NHE1 siRNA, HPASMCs, HPAECs, and HPAFs were cultured in a 2% oxygen chamber for 24 hours, and harvested for gene expression and cell counts to assay cell proliferation. *P < 0.05, compared with control group (n = 9 for each group).

Article Snippet: Cells Human pulmonary artery smooth muscle cells (HPASMCs), human pulmonary artery endothelial cells (HPAECs; Lonza, Inc., Walkersville, MD), and human pulmonary artery fibroblasts (HPAFs; ScienCell Research Laboratories, Carlsbad, CA) were used in this study.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Isolation, Control, Transfection, Cell Culture, Gene Expression