Microscopy:Article Title: Engineering vascularized 3D tissues: An optimized seeding protocol for collagen-based scaffolds
Article Snippet: .. Reagents/tools , Cell line and medium • Primary human umbilical vein endothelial cells, HUVECs (ATCC, Manasses, VA, cat # PCS–100–010) • Vascular cell basal medium (ATCC, cat # PCS–100–030) supplemented with endothelial cell growth kit-VEGF (ATCC, No. PCS–100–041) and 1 % penicillin/streptomycin (ThermoFisher, cat # 15,140,122) Equipment • PHCbi CO 2 Incubator • Countess 3 Automated Cell Counter (ThermoFisher Scientific) • NuWind Multi-Application Centrifuge • Leica TCS SP5 confocal microscope Reagents • Dulbecco’s phosphate-buffered saline (DPBS, ThermoFisher Scientific, cat # 14,040,182) • Trypsin-EDTA (ATCC, cat # 30–2101) • Cubic collagen-based Spongostan Dental MS0005 (Ethicon, Johnson & Johnson) • 4 % paraformaldehyde (PFA, Boster, cat # AR1068) • Phosphate buffered saline (PBS, ThermoFisher Scientific, cat # 10,010,023) • 1 % bovine serum albumin (BSA, Biophoretics, cat # BCZ11920.1 ) • Phalloidin (FITC, ThermoFisher Scientific, cat # F432) • 4′,6-diamidino-2-phenylindole (DAPI, Invitrogen, cat # D1306) Software • LAS AF software • ImageJ Angiogenesis Analysis • GraphPad Prism 10.5.0 Plasticware • T75 cm 2 cell culture flasks (VWR, cat # 10,062–860) • 15 ml conical centrifuge tubes (Oxford Lab Products, cat # OCT-15B) • 0.65 mL microcentrifuge tubes (VWR, cat # 87,003–290) • 12-well plates (VWR, cat # 10,861–556) • Scalpel handle #3 • Sterile surgical carbon steel blades (P&P Medical Surgical, size 10) • Tweezers • 20 μL sterile pipette tips (Oxford Lab Products, cat # XR-20-SLF) • 200 μL sterile pipette tips (Oxford Lab Products, cat # XR-200-SLF) • 1000 μL sterile pipette tips (Oxford Lab Products, cat # XR-1000-SLF) • 20 μL pipette (Eppendorf, cat # ES-20F) • 200 μL pipette (Eppendorf, cat # ES-200F) • 1000 μL pipette (Eppendorf, cat # ES-1000F) • 5 mL serological pipettes (Oxford Lab Products, cat # OSC-5C) • 10 mL serological pipettes (Oxford Lab Products, cat # OSP-10C). .. Experimental design , Endothelial cells were seeded in a commercial 3D collagen scaffold to achieve a vascular network. Scaffolded cells were dyed with DAPI and FITC to visualize branching, which was further quantified using ImageJ Angiogenesis Analysis. Scaffold saturation, cell density, one-sided and two-sided seeding, and incubation period were altered to determine the optimal method for achieving maximal vascularization and branching..
Saline:Article Title: Engineering vascularized 3D tissues: An optimized seeding protocol for collagen-based scaffolds
Article Snippet: .. Reagents/tools , Cell line and medium • Primary human umbilical vein endothelial cells, HUVECs (ATCC, Manasses, VA, cat # PCS–100–010) • Vascular cell basal medium (ATCC, cat # PCS–100–030) supplemented with endothelial cell growth kit-VEGF (ATCC, No. PCS–100–041) and 1 % penicillin/streptomycin (ThermoFisher, cat # 15,140,122) Equipment • PHCbi CO 2 Incubator • Countess 3 Automated Cell Counter (ThermoFisher Scientific) • NuWind Multi-Application Centrifuge • Leica TCS SP5 confocal microscope Reagents • Dulbecco’s phosphate-buffered saline (DPBS, ThermoFisher Scientific, cat # 14,040,182) • Trypsin-EDTA (ATCC, cat # 30–2101) • Cubic collagen-based Spongostan Dental MS0005 (Ethicon, Johnson & Johnson) • 4 % paraformaldehyde (PFA, Boster, cat # AR1068) • Phosphate buffered saline (PBS, ThermoFisher Scientific, cat # 10,010,023) • 1 % bovine serum albumin (BSA, Biophoretics, cat # BCZ11920.1 ) • Phalloidin (FITC, ThermoFisher Scientific, cat # F432) • 4′,6-diamidino-2-phenylindole (DAPI, Invitrogen, cat # D1306) Software • LAS AF software • ImageJ Angiogenesis Analysis • GraphPad Prism 10.5.0 Plasticware • T75 cm 2 cell culture flasks (VWR, cat # 10,062–860) • 15 ml conical centrifuge tubes (Oxford Lab Products, cat # OCT-15B) • 0.65 mL microcentrifuge tubes (VWR, cat # 87,003–290) • 12-well plates (VWR, cat # 10,861–556) • Scalpel handle #3 • Sterile surgical carbon steel blades (P&P Medical Surgical, size 10) • Tweezers • 20 μL sterile pipette tips (Oxford Lab Products, cat # XR-20-SLF) • 200 μL sterile pipette tips (Oxford Lab Products, cat # XR-200-SLF) • 1000 μL sterile pipette tips (Oxford Lab Products, cat # XR-1000-SLF) • 20 μL pipette (Eppendorf, cat # ES-20F) • 200 μL pipette (Eppendorf, cat # ES-200F) • 1000 μL pipette (Eppendorf, cat # ES-1000F) • 5 mL serological pipettes (Oxford Lab Products, cat # OSC-5C) • 10 mL serological pipettes (Oxford Lab Products, cat # OSP-10C). .. Experimental design , Endothelial cells were seeded in a commercial 3D collagen scaffold to achieve a vascular network. Scaffolded cells were dyed with DAPI and FITC to visualize branching, which was further quantified using ImageJ Angiogenesis Analysis. Scaffold saturation, cell density, one-sided and two-sided seeding, and incubation period were altered to determine the optimal method for achieving maximal vascularization and branching..
Software:Article Title: Engineering vascularized 3D tissues: An optimized seeding protocol for collagen-based scaffolds
Article Snippet: .. Reagents/tools , Cell line and medium • Primary human umbilical vein endothelial cells, HUVECs (ATCC, Manasses, VA, cat # PCS–100–010) • Vascular cell basal medium (ATCC, cat # PCS–100–030) supplemented with endothelial cell growth kit-VEGF (ATCC, No. PCS–100–041) and 1 % penicillin/streptomycin (ThermoFisher, cat # 15,140,122) Equipment • PHCbi CO 2 Incubator • Countess 3 Automated Cell Counter (ThermoFisher Scientific) • NuWind Multi-Application Centrifuge • Leica TCS SP5 confocal microscope Reagents • Dulbecco’s phosphate-buffered saline (DPBS, ThermoFisher Scientific, cat # 14,040,182) • Trypsin-EDTA (ATCC, cat # 30–2101) • Cubic collagen-based Spongostan Dental MS0005 (Ethicon, Johnson & Johnson) • 4 % paraformaldehyde (PFA, Boster, cat # AR1068) • Phosphate buffered saline (PBS, ThermoFisher Scientific, cat # 10,010,023) • 1 % bovine serum albumin (BSA, Biophoretics, cat # BCZ11920.1 ) • Phalloidin (FITC, ThermoFisher Scientific, cat # F432) • 4′,6-diamidino-2-phenylindole (DAPI, Invitrogen, cat # D1306) Software • LAS AF software • ImageJ Angiogenesis Analysis • GraphPad Prism 10.5.0 Plasticware • T75 cm 2 cell culture flasks (VWR, cat # 10,062–860) • 15 ml conical centrifuge tubes (Oxford Lab Products, cat # OCT-15B) • 0.65 mL microcentrifuge tubes (VWR, cat # 87,003–290) • 12-well plates (VWR, cat # 10,861–556) • Scalpel handle #3 • Sterile surgical carbon steel blades (P&P Medical Surgical, size 10) • Tweezers • 20 μL sterile pipette tips (Oxford Lab Products, cat # XR-20-SLF) • 200 μL sterile pipette tips (Oxford Lab Products, cat # XR-200-SLF) • 1000 μL sterile pipette tips (Oxford Lab Products, cat # XR-1000-SLF) • 20 μL pipette (Eppendorf, cat # ES-20F) • 200 μL pipette (Eppendorf, cat # ES-200F) • 1000 μL pipette (Eppendorf, cat # ES-1000F) • 5 mL serological pipettes (Oxford Lab Products, cat # OSC-5C) • 10 mL serological pipettes (Oxford Lab Products, cat # OSP-10C). .. Experimental design , Endothelial cells were seeded in a commercial 3D collagen scaffold to achieve a vascular network. Scaffolded cells were dyed with DAPI and FITC to visualize branching, which was further quantified using ImageJ Angiogenesis Analysis. Scaffold saturation, cell density, one-sided and two-sided seeding, and incubation period were altered to determine the optimal method for achieving maximal vascularization and branching..
Cell Culture:Article Title: Engineering vascularized 3D tissues: An optimized seeding protocol for collagen-based scaffolds
Article Snippet: .. Reagents/tools , Cell line and medium • Primary human umbilical vein endothelial cells, HUVECs (ATCC, Manasses, VA, cat # PCS–100–010) • Vascular cell basal medium (ATCC, cat # PCS–100–030) supplemented with endothelial cell growth kit-VEGF (ATCC, No. PCS–100–041) and 1 % penicillin/streptomycin (ThermoFisher, cat # 15,140,122) Equipment • PHCbi CO 2 Incubator • Countess 3 Automated Cell Counter (ThermoFisher Scientific) • NuWind Multi-Application Centrifuge • Leica TCS SP5 confocal microscope Reagents • Dulbecco’s phosphate-buffered saline (DPBS, ThermoFisher Scientific, cat # 14,040,182) • Trypsin-EDTA (ATCC, cat # 30–2101) • Cubic collagen-based Spongostan Dental MS0005 (Ethicon, Johnson & Johnson) • 4 % paraformaldehyde (PFA, Boster, cat # AR1068) • Phosphate buffered saline (PBS, ThermoFisher Scientific, cat # 10,010,023) • 1 % bovine serum albumin (BSA, Biophoretics, cat # BCZ11920.1 ) • Phalloidin (FITC, ThermoFisher Scientific, cat # F432) • 4′,6-diamidino-2-phenylindole (DAPI, Invitrogen, cat # D1306) Software • LAS AF software • ImageJ Angiogenesis Analysis • GraphPad Prism 10.5.0 Plasticware • T75 cm 2 cell culture flasks (VWR, cat # 10,062–860) • 15 ml conical centrifuge tubes (Oxford Lab Products, cat # OCT-15B) • 0.65 mL microcentrifuge tubes (VWR, cat # 87,003–290) • 12-well plates (VWR, cat # 10,861–556) • Scalpel handle #3 • Sterile surgical carbon steel blades (P&P Medical Surgical, size 10) • Tweezers • 20 μL sterile pipette tips (Oxford Lab Products, cat # XR-20-SLF) • 200 μL sterile pipette tips (Oxford Lab Products, cat # XR-200-SLF) • 1000 μL sterile pipette tips (Oxford Lab Products, cat # XR-1000-SLF) • 20 μL pipette (Eppendorf, cat # ES-20F) • 200 μL pipette (Eppendorf, cat # ES-200F) • 1000 μL pipette (Eppendorf, cat # ES-1000F) • 5 mL serological pipettes (Oxford Lab Products, cat # OSC-5C) • 10 mL serological pipettes (Oxford Lab Products, cat # OSP-10C). .. Experimental design , Endothelial cells were seeded in a commercial 3D collagen scaffold to achieve a vascular network. Scaffolded cells were dyed with DAPI and FITC to visualize branching, which was further quantified using ImageJ Angiogenesis Analysis. Scaffold saturation, cell density, one-sided and two-sided seeding, and incubation period were altered to determine the optimal method for achieving maximal vascularization and branching..
Sterility:Article Title: Engineering vascularized 3D tissues: An optimized seeding protocol for collagen-based scaffolds
Article Snippet: .. Reagents/tools , Cell line and medium • Primary human umbilical vein endothelial cells, HUVECs (ATCC, Manasses, VA, cat # PCS–100–010) • Vascular cell basal medium (ATCC, cat # PCS–100–030) supplemented with endothelial cell growth kit-VEGF (ATCC, No. PCS–100–041) and 1 % penicillin/streptomycin (ThermoFisher, cat # 15,140,122) Equipment • PHCbi CO 2 Incubator • Countess 3 Automated Cell Counter (ThermoFisher Scientific) • NuWind Multi-Application Centrifuge • Leica TCS SP5 confocal microscope Reagents • Dulbecco’s phosphate-buffered saline (DPBS, ThermoFisher Scientific, cat # 14,040,182) • Trypsin-EDTA (ATCC, cat # 30–2101) • Cubic collagen-based Spongostan Dental MS0005 (Ethicon, Johnson & Johnson) • 4 % paraformaldehyde (PFA, Boster, cat # AR1068) • Phosphate buffered saline (PBS, ThermoFisher Scientific, cat # 10,010,023) • 1 % bovine serum albumin (BSA, Biophoretics, cat # BCZ11920.1 ) • Phalloidin (FITC, ThermoFisher Scientific, cat # F432) • 4′,6-diamidino-2-phenylindole (DAPI, Invitrogen, cat # D1306) Software • LAS AF software • ImageJ Angiogenesis Analysis • GraphPad Prism 10.5.0 Plasticware • T75 cm 2 cell culture flasks (VWR, cat # 10,062–860) • 15 ml conical centrifuge tubes (Oxford Lab Products, cat # OCT-15B) • 0.65 mL microcentrifuge tubes (VWR, cat # 87,003–290) • 12-well plates (VWR, cat # 10,861–556) • Scalpel handle #3 • Sterile surgical carbon steel blades (P&P Medical Surgical, size 10) • Tweezers • 20 μL sterile pipette tips (Oxford Lab Products, cat # XR-20-SLF) • 200 μL sterile pipette tips (Oxford Lab Products, cat # XR-200-SLF) • 1000 μL sterile pipette tips (Oxford Lab Products, cat # XR-1000-SLF) • 20 μL pipette (Eppendorf, cat # ES-20F) • 200 μL pipette (Eppendorf, cat # ES-200F) • 1000 μL pipette (Eppendorf, cat # ES-1000F) • 5 mL serological pipettes (Oxford Lab Products, cat # OSC-5C) • 10 mL serological pipettes (Oxford Lab Products, cat # OSP-10C). .. Experimental design , Endothelial cells were seeded in a commercial 3D collagen scaffold to achieve a vascular network. Scaffolded cells were dyed with DAPI and FITC to visualize branching, which was further quantified using ImageJ Angiogenesis Analysis. Scaffold saturation, cell density, one-sided and two-sided seeding, and incubation period were altered to determine the optimal method for achieving maximal vascularization and branching..
Transferring:Article Title: Engineering vascularized 3D tissues: An optimized seeding protocol for collagen-based scaffolds
Article Snippet: .. Reagents/tools , Cell line and medium • Primary human umbilical vein endothelial cells, HUVECs (ATCC, Manasses, VA, cat # PCS–100–010) • Vascular cell basal medium (ATCC, cat # PCS–100–030) supplemented with endothelial cell growth kit-VEGF (ATCC, No. PCS–100–041) and 1 % penicillin/streptomycin (ThermoFisher, cat # 15,140,122) Equipment • PHCbi CO 2 Incubator • Countess 3 Automated Cell Counter (ThermoFisher Scientific) • NuWind Multi-Application Centrifuge • Leica TCS SP5 confocal microscope Reagents • Dulbecco’s phosphate-buffered saline (DPBS, ThermoFisher Scientific, cat # 14,040,182) • Trypsin-EDTA (ATCC, cat # 30–2101) • Cubic collagen-based Spongostan Dental MS0005 (Ethicon, Johnson & Johnson) • 4 % paraformaldehyde (PFA, Boster, cat # AR1068) • Phosphate buffered saline (PBS, ThermoFisher Scientific, cat # 10,010,023) • 1 % bovine serum albumin (BSA, Biophoretics, cat # BCZ11920.1 ) • Phalloidin (FITC, ThermoFisher Scientific, cat # F432) • 4′,6-diamidino-2-phenylindole (DAPI, Invitrogen, cat # D1306) Software • LAS AF software • ImageJ Angiogenesis Analysis • GraphPad Prism 10.5.0 Plasticware • T75 cm 2 cell culture flasks (VWR, cat # 10,062–860) • 15 ml conical centrifuge tubes (Oxford Lab Products, cat # OCT-15B) • 0.65 mL microcentrifuge tubes (VWR, cat # 87,003–290) • 12-well plates (VWR, cat # 10,861–556) • Scalpel handle #3 • Sterile surgical carbon steel blades (P&P Medical Surgical, size 10) • Tweezers • 20 μL sterile pipette tips (Oxford Lab Products, cat # XR-20-SLF) • 200 μL sterile pipette tips (Oxford Lab Products, cat # XR-200-SLF) • 1000 μL sterile pipette tips (Oxford Lab Products, cat # XR-1000-SLF) • 20 μL pipette (Eppendorf, cat # ES-20F) • 200 μL pipette (Eppendorf, cat # ES-200F) • 1000 μL pipette (Eppendorf, cat # ES-1000F) • 5 mL serological pipettes (Oxford Lab Products, cat # OSC-5C) • 10 mL serological pipettes (Oxford Lab Products, cat # OSP-10C). .. Experimental design , Endothelial cells were seeded in a commercial 3D collagen scaffold to achieve a vascular network. Scaffolded cells were dyed with DAPI and FITC to visualize branching, which was further quantified using ImageJ Angiogenesis Analysis. Scaffold saturation, cell density, one-sided and two-sided seeding, and incubation period were altered to determine the optimal method for achieving maximal vascularization and branching..
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