At indicated time points after differentiation, h-iECs were dissociated into single cells and sorted into CD31+ and CD31− cells using magnetic beads coated with anti-human CD31 antibodies (DynaBeads, Thermo Fisher Scientific, catalog no. 11155D). The purified CD31+ h-iECs were then expanded in culture on 10-cm dishes coated with 1% gelatin. Culture medium for h-iECs was prepared by adding Endothelial Cell Growth medium 2 kit supplements into basal medium (except for hydrocortisone; PromoCell, catalog no. C22111) with 1× GlutaMax supplement and 10 μM SB431542.
C 22111
Manufactured by PromoCell
Sourced in Germany
The C-22111 is a laboratory centrifuge that can be used for a variety of applications requiring the separation of materials by density or size. It features a brushless motor and a digital display for setting speed and time. The centrifuge has a maximum speed of 6,000 rpm and can accommodate a variety of tube sizes and rotor configurations.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Dynabead, by Thermo Fisher Scientific (1 mentions)
C 22062, by PromoCell (1 mentions)
C 12205, by PromoCell (1 mentions)
C 12511, by PromoCell (1 mentions)
M200500, by Thermo Fisher Scientific (1 mentions)
Af 100 15, by Merck Group (1 mentions)
H3149 50ku, by Merck Group (1 mentions)
Fb 01, by Omega Scientific (1 mentions)
C 12212, by PromoCell (1 mentions)
Hdmec, by Lonza (1 mentions)
Egm 2, by PromoCell (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Huvecs, by Lonza (1 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
Dulbecco modified eagle medium (dmem), by Biowest (1 mentions)
6 protocols using c 22111
1
Isolation and Expansion of h-iECs
2
Vesseloid Formation from HUVECs and SMCs
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HUVECs and SMCs (C-12205 and C-12511, PromoCell) were maintained in endothelial growth medium 2 (EGM2; C-22111, PromoCell) and SMC growth medium (SMCGM2; C-22062, PromoCell), respectively, under water-saturated 5% CO2 atmosphere at 37°C. Cells were grown in collagen-I–coated flasks (10190103, Thermo Fisher Scientific) and detached at passage 5 for vesseloid formation.
3
Culturing Immortalized HUVECs and Myeloid Cell Lines
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Immortalized HUVECs (E4EC) 13 were obtained from S. Rafii's laboratory and were cultured on gelatin-coated flasks with Medium 199 (GE Life Sciences, sh30253.01) containing 20% FBS (FB-01, Omega Scientific), 1% penicillin-streptomycin, 1% l-glutamine, 10 mM HEPES (Thermo Fisher Scientific, 15140-122, 2503-081 and 15630080), human FGF (20 ng ml -1 ) (R&D Systems, 233-FB), human EGF (10 ng ml -1 ), human IGF-I (10 ng ml -1 ) (Peprotech, AF-100-15 and 100-11) and heparin (50 μg ml -1 ) (Sigma-Aldrich, H3149-50KU). HUVECs (Thermo Fisher Scientific, C0035C) were cultured with Medium 200 supplemented with low-serum growth supplement (Thermo Fisher Scientific, M200500 and S00310) or a endothelial growth medium 2 kit with the provided supplements (EGF, FGF, IGF, but no VEGF) (PromoCell, C-22111). KG1 HSC-like AML cells (obtained from Chute's Laboratory, originally from the American Type Culture Collection) and MKPL1 cells (obtained from K. Li, originally from DSMZ) were cultured in RPMI with 10% FBS, 1% penicillinstreptomycin and 1% l-glutamine. Cell lines were not authenticated and not tested for mycoplasma contamination.
4
Isolation and Culture of HUVECs, HUASMCs
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Biomaterials Science from the endothelium of the veins or arteries of human umbilical cords after informed consent approved by the Ethics Committee of the Faculty of Medicine, RWTH Aachen University (EK 424/19). HUVECs were cultivated in T75 cell culture flasks previously coated with 2% (w/v) gelatin, whereas HUASMCs were cultured in uncoated flasks. As growth medium, endothelial cell basal medium (C-22111, PromoCell, Heidelberg, Germany) for HUVECs and 1× DMEM (21885, ThermoFisher Scientific, Waltham, MA, USA) for HUASMCs were used and cells were used for experiments between passage two and five.
5
Cultivation of Human Dermal Microvascular Endothelial Cells
6
Diverse Cell Culture Protocols
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GFP+ MDA-MB231 cells were kindly provided by Dr. Julia Steitz, Institute of Laboratory Animal Science, RWTH University Hospital, Aachen, Germany. The cells were cultured in DMEM (11965092, Thermofisher Scientific) with 10% fetal bovine serum (FBS). Osteoblasts were procured from Sigma Aldrich (406-05F), and cultured in Osteoblast growth media (417–500, Sigma Aldrich). Fibroblasts were obtained by isolation of skin biopsies that were surgically extracted from healthy volunteers (Department of Dermatology, RWTH Aachen University Hospital, Germany). The extraction and isolation were conducted in accordance with the Declaration of Helsinki principles and was approved by the ethical committee of RWTH Aachen University Hospital, Germany, Project Number EK188/4. Fibroblasts were cultured in DMEM with 10% FBS. HUVECs (Lonza, Passage < 4) were cultured in an endothelial growth medium (C-22111, Promocell). For the culture of VBCTs, DMEM: EGM (1:1) with 10% fetal bovine serum (FBS, Biowest) and 50 µg/ml Ascorbic acid (A92902, Sigma) was used.
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