primary bronchial smooth muscle cells bsmcs  (PromoCell)

Journal: Nature Communications

Article Title: SARS-CoV-2 infection of human pluripotent stem cell-derived vascular cells reveals smooth muscle cells as key mediators of vascular pathology during infection

doi: 10.1038/s41467-024-54917-4

Figure Lengend Snippet: A Schematic of directed differentiation of hPSCs to vascular endothelial cells (ECs), smooth muscle cells (SMCs), and pericytes (PCs) (Created using BioRender). B Bulk RNA sequencing was performed on hPSC-derived ECs, PCs, and SMCs. The normalized expression values of known EC, PC, or SMC marker genes were quantitated over three biological replicates. The values plotted represent scaled normalized expression values for each gene across samples ( C ) (left) Principal component analysis (PCA) on bulk-RNA sequencing data from hPSCs, hPSC-ECs, hPSC-PCs, and hPSC-SMCs and primary endothelial cells (HUVECs), primary pericytes, and primary bronchial smooth muscle cells (BSMCs). (right) Mural cell-only PCA analysis (Primary pericytes, BSMCs, hPSC-PCs, and hPSC-SMCs) ( D ) Expression of PECAM1 and VE-Cadherin was quantitated by flow cytometry on hPSCs or on two independent differentiations of ECs. Data points on the bar graph represent values from two independent differentiations. Expression of VE-Cadherin, VWF, and PECAM1 in hPSC-derived ECs was observed by immunofluorescence. E Expression of NG2 was quantitated by flow cytometry on hPSCs or on two independent differentiations of PCs. Data points on bar graph represent values from two independent differentiations. Expression of PDGFR-B, and PDGFR-A was quantitated by flow cytometry on HEK293 cells, human dermal fibroblast (hDF) and two independent PC differentiation. Expression of SMA, PDGFR-B, and NG2 in hPSC derived PCs was observed by immunofluorescence. F Expression of PDGFR-B or SMA was quantitated by flow cytometry on hPSCs or on two independent differentiations of SMCs. Data points on the bar graph represent values from two independent differentiations. Expression of SMA, PDGFR-B, and NG2 in hPSC-derived SMCs was observed by immunofluorescence. Scale bar = 50 µm for all immunofluorescence images.

Article Snippet: Primary bronchial smooth muscle cells (BSMCs) were obtained from PromoCell (Cat# C12561).

Techniques: RNA Sequencing Assay, Derivative Assay, Expressing, Marker, Flow Cytometry, Immunofluorescence

Journal: Respiratory Research

Article Title: Differential regulation of neurotrophin expression in human bronchial smooth muscle cells

doi: 10.1186/1465-9921-7-18

Figure Lengend Snippet: Constitutive expression of neurotrophins by non-treated HBSMC evaluated by RT-PCR and ELISA. (A) Expression of NGF, BDNF, NT-3 mRNA and 18S rRNA. Reversed transcribed (RT+) and non-reversed transcribed (RT-) mRNA or rRNA are displayed. (B) NGF (white bars) and BDNF (black bars) protein secretion determined by ELISA. Data presented as mean ± SEM of 3–4 independent experiments performed in duplicate. In (B) ***: p < 0.001 for NGF, and #: p < 0.05 and ##: p < 0.01 for BDNF, versus 2.5 h

Article Snippet: HBSMC in primary culture from a healthy donor (Promocell, Heidelberg, Germany) were grown in monolayer in DMEM (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% FBS (Invitrogen, Rockville, MD, USA), 100 U/mL penicillin, 100 μg/mL streptomycin, 2 μg/mL amphotericin B (Fungizone ® , all Sigma-Aldrich) and 0.12 IU/mL insulin (Lilly, St Cloud, France) in 25 cm 2 flasks (Becton Dickinson Falcon, Franklin Lakes, NJ, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: Differential regulation of neurotrophin expression in human bronchial smooth muscle cells

doi: 10.1186/1465-9921-7-18

Figure Lengend Snippet: Time-course effects of IL-1β (10 U/mL) on NGF and BDNF mRNA and protein expression in HBSMC evaluated with real-time RT-PCR and ELISA, respectively. (A) NGF mRNA; (B) NGF protein; (C) BDNF mRNA and (D) BDNF protein. The IL-1β-dependent effects on neurotrophin mRNA expression and secreted protein are expressed as change in neurotrophin mRNA (%) and protein release (pg/mL) from baseline level of time-related control. Unstimulated control is set to 0 (% or pg/mL, respectively). Data are presented as mean ± SEM of 4–10 independent experiments performed in duplicate. In (A) and (C): **: p < 0.01 versus 0.5 h, ***: p < 0.001 versus 0.5 h. In (B) and (D): ***: p < 0.001 versus 2.5 h.

Article Snippet: HBSMC in primary culture from a healthy donor (Promocell, Heidelberg, Germany) were grown in monolayer in DMEM (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% FBS (Invitrogen, Rockville, MD, USA), 100 U/mL penicillin, 100 μg/mL streptomycin, 2 μg/mL amphotericin B (Fungizone ® , all Sigma-Aldrich) and 0.12 IU/mL insulin (Lilly, St Cloud, France) in 25 cm 2 flasks (Becton Dickinson Falcon, Franklin Lakes, NJ, USA).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: Differential regulation of neurotrophin expression in human bronchial smooth muscle cells

doi: 10.1186/1465-9921-7-18

Figure Lengend Snippet: Dose-dependent effects of IL-1β on neurotrophin mRNA expression in HBSMC evaluated by real-time RT-PCR. NGF mRNA expression evaluated at 6 h (solid squares); BDNF mRNA expression evaluated at 48 h (open triangle); NT-3 mRNA evaluated at 48 h (closed triangles). The IL-1β-dependent effects on neurotrophin mRNA expression are expressed as change in neurotrophin mRNA (%) from baseline level of time-related control. Unstimulated control is set to 0 %. Data are expressed as mean ± SEM of 3 independent experiments performed in duplicate. *: p < 0.05 for NGF and #: p < 0.05 for BDNF versus unstimulated control.

Article Snippet: HBSMC in primary culture from a healthy donor (Promocell, Heidelberg, Germany) were grown in monolayer in DMEM (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% FBS (Invitrogen, Rockville, MD, USA), 100 U/mL penicillin, 100 μg/mL streptomycin, 2 μg/mL amphotericin B (Fungizone ® , all Sigma-Aldrich) and 0.12 IU/mL insulin (Lilly, St Cloud, France) in 25 cm 2 flasks (Becton Dickinson Falcon, Franklin Lakes, NJ, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Respiratory Research

Article Title: Differential regulation of neurotrophin expression in human bronchial smooth muscle cells

doi: 10.1186/1465-9921-7-18

Figure Lengend Snippet: Effects of the COX-inhibitors indomethacin and NS-398 (both 10 μM) on IL-1β (10 U/mL)-stimulated NGF (A) and BDNF (B) secretion by HBSMC, evaluated by ELISA. Data are presented as mean ± SEM of 6 independent experiments. ***: p < 0.001 versus control, and #: p < 0.05, ##: p < 0.01 versus IL-1β (10 U/mL) alone.

Article Snippet: HBSMC in primary culture from a healthy donor (Promocell, Heidelberg, Germany) were grown in monolayer in DMEM (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% FBS (Invitrogen, Rockville, MD, USA), 100 U/mL penicillin, 100 μg/mL streptomycin, 2 μg/mL amphotericin B (Fungizone ® , all Sigma-Aldrich) and 0.12 IU/mL insulin (Lilly, St Cloud, France) in 25 cm 2 flasks (Becton Dickinson Falcon, Franklin Lakes, NJ, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: Differential regulation of neurotrophin expression in human bronchial smooth muscle cells

doi: 10.1186/1465-9921-7-18

Figure Lengend Snippet: Time-course effects of IFN-γ (10 ng/mL) on NGF (A) and BDNF (B) mRNA expression in HBSMC evaluated by real-time RT-PCR. The IFN-γ-dependent effects on neurotrophin mRNA expression are expressed as change in neurotrophin mRNA (%) from baseline level of time-related control. Unstimulated control for each time-point is set to 0 %. Data are presented as mean ± SEM of 6 independent experiments performed in duplicate. *: p < 0.05, **: p < 0.01, ***: p > 0.001 versus 0.5 h.

Article Snippet: HBSMC in primary culture from a healthy donor (Promocell, Heidelberg, Germany) were grown in monolayer in DMEM (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% FBS (Invitrogen, Rockville, MD, USA), 100 U/mL penicillin, 100 μg/mL streptomycin, 2 μg/mL amphotericin B (Fungizone ® , all Sigma-Aldrich) and 0.12 IU/mL insulin (Lilly, St Cloud, France) in 25 cm 2 flasks (Becton Dickinson Falcon, Franklin Lakes, NJ, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Respiratory Research

Article Title: Differential regulation of neurotrophin expression in human bronchial smooth muscle cells

doi: 10.1186/1465-9921-7-18

Figure Lengend Snippet: Dose-dependent effects of IFN-γ and IL-4 on IL-1β (10 U/mL)-stimulated NGF expression in HBSMC evaluated by real-time RT-PCR. Solid triangles illustrate effects of IFN-γ (A) and IL-4 (B) alone and solid squares illustrate IL-1β in combination with IFN-γ (A) and IL-4 (B). The IFN-γ- or IL-4-dependent effects on IL-1β-stimulated NGF mRNA expression are expressed as change in NGF mRNA (%) from baseline level of time-related control. Unstimulated control is set to 0 %. Data are presented as mean ± SEM of 3–4 independent experiments performed in duplicate. *: p < 0.05, **: p < 0.01, ***: p < 0.001 versus IL-1β (10 U/mL) alone.

Article Snippet: HBSMC in primary culture from a healthy donor (Promocell, Heidelberg, Germany) were grown in monolayer in DMEM (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% FBS (Invitrogen, Rockville, MD, USA), 100 U/mL penicillin, 100 μg/mL streptomycin, 2 μg/mL amphotericin B (Fungizone ® , all Sigma-Aldrich) and 0.12 IU/mL insulin (Lilly, St Cloud, France) in 25 cm 2 flasks (Becton Dickinson Falcon, Franklin Lakes, NJ, USA).

Techniques: Expressing, Quantitative RT-PCR