Cells (HPMEC) were obtained from Promocell (C-12281). They were cultured on coated flasks (Angioproteomie, cAP-01) before being plated for co-culture on the microfluidic platform. Prior to experiment, cells were given fresh media (Lifeline Technology, LM-0002) every other day. The cells were dissociated from the sub-culture flasks using 0.05% trypsin (ThermoFisher Scientific, 25300).
C 12281
Manufactured by PromoCell
Sourced in Germany
C-12281 is a laboratory centrifuge designed for general-purpose applications. It features a compact and lightweight design, providing a convenient solution for sample separation and preparation in various laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
C 22120, by PromoCell (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Cap 01, by Angio-Proteomie (1 mentions)
C 22022, by PromoCell (1 mentions)
Cc 3162, by Lonza (1 mentions)
Axiovert 200m inverted microscope, by Zeiss (1 mentions)
C 12212, by PromoCell (1 mentions)
Pcs 100 011, by American Type Culture Collection (1 mentions)
Cc 2545, by Lonza (1 mentions)
C 12203, by PromoCell (1 mentions)
Fetal bovine serum (fbs), by Omega Scientific (1 mentions)
Cc 3182, by Lonza (1 mentions)
Cc 2581, by Lonza (1 mentions)
G9391, by Merck Group (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
7 protocols using c 12281
1
Cultivation of Pulmonary Endothelial Cells
2
Culturing Primary Human Pulmonary Endothelial Cells
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Primary human pulmonary microvascular endothelial cells (HPMECs) derived from a male donor (C-12281; PromoCell) were maintained in endothelial cell basal medium (BM) MV2 supplemented with 5% Fetal calf serum (FCS), ascorbic acid and hydrocortisone, together with the following growth factors: human epidermal growth factor, human fibroblast growth factor, insulin-like growth factor-1 and human vascular endothelial growth factor (VEGF), according to the manufacturer’s recommendations. Cells were used for experiments between passages four and six.
3
Isolation and Culture of HLMVEC
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HLMVEC were obtained from small vessels within normal lung tissue (Promocell,C-12281 Germany, www.promocell.com ). HLMVEC were cultured in endothelial cell growth medium (EBM2 basal medium supplemented with human epidermal growth factor, vascular endothelial growth factor, R3-insulin-like growth factor-1, ascorbic acid, hydrocortisone, human fibroblast growth factor-beta, 5% fetal bovine serum, gentamicin/amphotericin-B and 1% penicillin/streptomycin [Promocell, C-22022,Germany, www.promocell.com ]) and incubated in a humidified incubator with 5% CO2 at 37 °C. Change the growth medium 24 h after inoculation, and every other day thereafter. When the cells reach 70–85% confluence, they are subcultured. HLMVEC with a total passage number of < 9 were used in all experiments.
4
Culturing Primary Human Lung Endothelial Cells
5
Endothelial Cell Culture and Characterization
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Human ECs were purchased from different vendors to capture a broad spectrum of sampling. Specifically, we obtained HUVECs (C2519A; Lonza; C-12203; PromoCell), HSaVECs (HSVEC/A; VEC-Technologies; cAP-0019; Angio-Proteomie), HAECs (PCS-100-011; ATCC; 6100; ScienCell), HIAECs (CC-2545; Lonza; cAP-0020; Angio-Proteomie), HBMVECs (ACBRI 376 V; Cell-Systems; cAP-0002; Angio-Proteomie), HUMVECs (C-12295; PromoCell; 7000; ScienCell), HLMVECs (3000; ScienCell; C-12281; PromoCell), HAMVECs (7200; ScienCell), and HDMECs (C-12212; PromoCell; 2000; ScienCell). All types of ECs were cultured in EGM-2 endothelial growth medium (CC-3162; Lonza) supplemented with 5% FBS (Omega Scientific) and were maintained at 37°C with 5% CO2. Similar EC passages 4–6 were used for final experiments. Human dermal fibroblasts were a gift from W. Lowry (University of California, Los Angeles, Los Angeles, CA), expanded in DMEM supplemented with 10% FBS, and cultured in EMG-2 medium over 48 h before RNA expression experiments. Brightfield images were acquired by using an Axiovert 200M inverted microscope (Zeiss).
6
Characterization of Human Lung Cell Types
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Lung pericytes were maintained in Pericyte medium (sc-1201, Provitro) supplemented with growth factors and 2% fetal bovine serum. All experiments were carried out between passages 7-11 from a minimum of three subjects.
Each experiment was repeated a minimum of three times, and each time, cells from different human subjects were used to attest for biological heterogeneity. For functional experiments (BrdU incorporation Assay, Wound healing assay), minimum three technical replicas were used within one experiment.
Human pulmonary artery smooth muscle cells (PASMCs) were purchased from Lonza (CC-2581) and cultured in smooth muscle cell growth medium (CC-3182, Lonza) according to the protocol previously described 46 . All experiments were carried out at passage 6 from a minimum of three subjects.
Human microvascular lung endothelial cells (MLVECs) were purchased from Promocell (c-12281). The cells were maintained in microvascular endothelial medium MV (c-22120, Promocell) according to the manufacturer's instructions. RIA was carried out in passage 5.
Each experiment was repeated a minimum of three times, and each time, cells from different human subjects were used to attest for biological heterogeneity. For functional experiments (BrdU incorporation Assay, Wound healing assay), minimum three technical replicas were used within one experiment.
Human pulmonary artery smooth muscle cells (PASMCs) were purchased from Lonza (CC-2581) and cultured in smooth muscle cell growth medium (CC-3182, Lonza) according to the protocol previously described 46 . All experiments were carried out at passage 6 from a minimum of three subjects.
Human microvascular lung endothelial cells (MLVECs) were purchased from Promocell (c-12281). The cells were maintained in microvascular endothelial medium MV (c-22120, Promocell) according to the manufacturer's instructions. RIA was carried out in passage 5.
7
Primary HPMEC and NCI-H441 Cell Culture
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The primary human pulmonary microvascular
endothelial cells (HPMEC, Promocell, C-12281) were maintained and
cultured in 2 wt % gelatin-coated culture (Sigma-Aldrich, G9391) flasks
using microvascular endothelial growth media (EGM) (Promocell, C-22120).
Epithelial human lung adenocarcinoma cell line (NCI-H441, ATCC, ATCC-HTB-174)
was cultured in RPMI-1640 (Thermofisher Scientific, 21875) supplemented
with 1% penicillin/streptomycin (Thermofisher Scientific, 15140) and
10% fetal bovine serum (FBS, BH Biowest, 5181). The environmental
conditions for both cell types were 37 °C and 5% CO2. When cells achieved 85% confluency, they were either subcultured
or used for experiments. While HPMEC were not subcultured above passage
4, NCI-H441 cells were used until passage 20 for experiments.
endothelial cells (HPMEC, Promocell, C-12281) were maintained and
cultured in 2 wt % gelatin-coated culture (Sigma-Aldrich, G9391) flasks
using microvascular endothelial growth media (EGM) (Promocell, C-22120).
Epithelial human lung adenocarcinoma cell line (NCI-H441, ATCC, ATCC-HTB-174)
was cultured in RPMI-1640 (Thermofisher Scientific, 21875) supplemented
with 1% penicillin/streptomycin (Thermofisher Scientific, 15140) and
10% fetal bovine serum (FBS, BH Biowest, 5181). The environmental
conditions for both cell types were 37 °C and 5% CO2. When cells achieved 85% confluency, they were either subcultured
or used for experiments. While HPMEC were not subcultured above passage
4, NCI-H441 cells were used until passage 20 for experiments.
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