Human umbilical cords were obtained from Xinhua Hospital affiliated with Shanghai Jiao Tong University School of Medicine after obtaining informed consent from all participants and approval from the institutional ethical committee (approval number: XHEC‐C‐2020‐092‐1). Human umbilical cord mesenchymal stem cells (MSCs) were donated by the National Stem Cell Resource Centre, Beijing. The transduction method with red fluorescent protein (RFP) into HUVECs were as previously reported.17 RFP‐expressing HUVECs (RFP‐HUVECs) were cultured in EMG2 medium (complete EGM‐2 BulletKit™, Lonza). Green fluorescent protein‐expressing MSCs (GFP‐MSCs) were cultured in Dulbecco's modified Eagle medium containing high glucose and sodium pyruvate (DMEM, Gibco) supplemented with 15% fetal bovine serum (FBS, Bioind), 1% non‐essential amino acid solution (NEAA, Gibco), 1% GlutaMAX™ (Gibco), and 1% penicillin/streptomycin (Gibco). The culture medium was added and changed every 2 days for culturing at 37 °C with 5% CO2. RFP‐HUVECs and GFP‐MSCs were not used beyond the 10th passage.
Complete egm 2 bulletkit
Manufactured by Lonza
Sourced in United States
The Complete EGM‐2 BulletKit™ is a laboratory equipment product designed for cell culture applications. It provides a comprehensive set of components required to support the growth and maintenance of endothelial cells in vitro. The kit includes essential supplements, growth factors, and other necessary reagents to facilitate optimal cell culture conditions.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Haecs, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
Mem neaa, by Thermo Fisher Scientific (1 mentions)
Live dead viability kit, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde (pfa), by Beyotime (1 mentions)
Tb green premix ex taq, by Takara Bio (1 mentions)
Phosphate buffered saline (pbs), by Sartorius (1 mentions)
Rnaiso plus, by Takara Bio (1 mentions)
Resazurin, by Maclin Biochemical Technology (1 mentions)
3 protocols using complete egm 2 bulletkit
1
Umbilical Cord MSCs and RFP-HUVECs Protocol
2
Urolithins Effects on Human Endothelial Cells
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HAECs (purchased from GIBCO Life Technologies, Carlsbad, CA, USA) were cultured in complete EGM-2 Bullet Kit (Lonza, Basel, Switzerland) at 37 °C in a 5% CO2 humidified incubator and used at passages 3–7 in all the experiments. HAEC were seeded in T25 flasks at a density of 1 × 104 cells/cm2. On reaching 70%–80% confluence, cells were cultured overnight in serum free-media and then incubated with urolithins or with vehicle (0.1% DMSO as negative control) for 5 min and 24 h. Uro A, Uro B and Uro Bgluc were incubated at a concentration of 15 μM. A mixture of urolithins at 5 μM each was also added to the culture medium in order to test the combined effects of metabolites.
3
Alginate-Based 3D Bioprinting for Vascular Tissue
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The main reagents used in this study were: sodium alginate (Alg, from brown algea, low viscosity, A1112, Sigma), sodium alginate (from brown algea, medium viscosity, 71,238, Sigma), gelatin (Gel, from porcine skin, G1890, Sigma), calcium chloride (CaCl2, C5670, Sigma), glutamine transaminase (TG, G8661, Solarbio), phosphate buffered saline (PBS, 02-02402ACS, Biological Industries), endothelial cell growth medium-2 (complete EGM-2 BulletKit, CC3162, Lonza), fetal bovine serum (FBS, 10,099–141, Gibco), Dulbecco's modified eagle medium (DMEM, C11965500BT, Gibco), GlutaMAX (35,050,061, Thermo), minimum essential medium non-essential amino acids (MEM NEAA, 11,140,050, Gibco), penicillin-Streptomycin (SP, 60162ES76, Yeasen), trypsin-EDTA (25200072, Gibco), resazurin (R817239, Macklin), RNAiso Plus (9108, Takara), PrimeScript™ RT reagent Kit (RR047A, Takara), TB Green® Premix Ex Taq (RR420A, Takara), 4 % paraformaldehyde (P0099, Beyotime), bovineVKO9 serum albumin (BSA, SW3015, Solarbio), Triton X-100 (X100, Sigma), Live/Dead Viability Kit (L-3224, Life technologies). GelMA was prepared according to the method reported by Liu et al. [28 ]. PSCs were donated by Dong Qiu Group [29 ].
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