The study protocol was approved by the institutional review board of Duksung Women's University (IRB No. 2017-002-001). ECFCs were isolated from the adherent mononuclear cell (MNC) fraction of human peripheral blood using CD31-coated magnetic beads (Invitrogen, MA, USA) as described previously [10 (link)]. The isolated ECFCs were expanded on 1% gelatin-coated plates (BD Biosciences, NJ, USA) using endothelial growth medium-2 (EGM-2 without hydrocortisone; Lonza, MD, USA) supplemented with 10% fetal bovine serum (FBS; Atlas Biologicals, CO, USA) and 1% glutamine-penicillin-streptomycin (GPS; Gibco, MA, USA). ECFCs between passages 7 and 10 were used for all experiments.
Gelatin coated plates
Manufactured by BD
Sourced in United States
Gelatin-coated plates are laboratory equipment used for cell culture and microbiology applications. They provide a stable and uniform surface for cell attachment and growth. The gelatin coating enhances cell adhesion and promotes a suitable environment for cell culture experiments.
Automatically generated - may contain errors
Lab products found in correlation
Cd31 coated magnetic beads, by Thermo Fisher Scientific (4 mentions)
Msc growth medium, by Lonza (2 mentions)
Fetal bovine serum (fbs), by Atlas Biologicals (2 mentions)
Glutamine penicillin streptomycin gps, by Thermo Fisher Scientific (2 mentions)
Egm 2 without hydrocortisone, by Lonza (1 mentions)
Glutamine penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Egm mv 2 without hydrocortisone, by PromoCell (1 mentions)
Endothelial growth medium 2 egm 2, by Lonza (1 mentions)
4 protocols using gelatin coated plates
1
ECFC Isolation and Expansion Protocol
2
Isolation and Expansion of ECFCs and MSCs
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The study protocol was approved by the institutional review board of Duksung Women’s University (IRB No. 2017-002-01). The endothelial colony-forming cells (ECFCs) were isolated from human peripheral blood provided from one normal adult male donor. Some CD31-coated magnetic beads (Invitrogen, Waltham, MA, USA) were used to isolate ECFCs from the adherent mononuclear cell fraction of blood, as described in a previous report [19 (link)]. The isolated ECFCs were expanded on 1% gelatin-coated plates (BD Biosciences, Franklin Lakes, NJ, USA) using endothelial cell growth medium (EGM-2; Lonza, Basel, Switzerland) without hydrocortisone, supplemented with 10% fetal bovine serum (FBS; Atlas Biologicals, Fort Collins, CO, USA) and 1% glutamine–penicillin–streptomycin (GPS; Gibco, Waltham, MA, USA). Mesenchymal stem cells (MSCs) isolated from one normal human adult bone marrow were purchased from Lonza and then cultured using MSC growth medium (Lonza) supplemented with 10% FBS and 1% GPS. The ECFCs and MSCs between passages 7 and 10 were used for all experiments.
3
Isolation and Expansion of Endothelial Colony-Forming Cells
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The study protocol was approved by the Institutional Review Board (IRB) of Duksung Women's University (IRB No. 2017-002-001, 2018-007-006). ECFCs were isolated from the adherent mononuclear cell fraction of human peripheral blood using CD31-coated magnetic beads (Invitrogen, Waltham, MA, USA) as described previously.14 (link) Isolated ECFCs were expanded on 1% gelatin-coated plates (BD Biosciences, Franklin Lakes, NJ, USA) using EC growth medium MV 2 (EGM-MV 2 without hydrocortisone; Promocell, Heidelberg, Germany) supplemented with 10% fetal bovine serum (FBS; Atlas Biologicals, Fort Collins, CO, USA) and 1% glutamine-penicillin-streptomycin (GPS; Gibco, Waltham, MA, USA). ECFCs obtained between passages 7 and 10 were used for all experiments.
4
Isolation and Expansion of ECFCs and MSCs
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+ Expand
The study protocol was approved by the institutional review board of Duksung Women’s University (IRB No. 2017-002-001). Human peripheral blood was provided from the national biobank. ECFCs were isolated from the adherent mononuclear cell (MNC) fraction using CD31-coated magnetic beads (Invitrogen, MA, USA) as described in the previous report (Melero-Martin et al., 2008 (link)). The isolated ECFCs were expanded on 1% gelatin-coated plates (BD Biosciences, NJ, USA) using endothelial growth medium-2 (EGM-2; Lonza, MD, USA) without hydrocortisone supplemented with 10% fetal bovine serum (FBS; Atlas Biologicals, CO, USA) and 1% glutamine-penicillin-streptomycin (GPS; Gibco, MA, USA). In all experiments, ECFCs from passages 7 to 10 were used.
MSCs were obtained from the MNC fraction of human adult bone marrow (Lonza). MSCs were cultured in MSC growth medium (Lonza) containing 10% FBS and 1% GPS until 80% confluence was achieved. MSCs from passage numbers 5 to 8 were used.
MSCs were obtained from the MNC fraction of human adult bone marrow (Lonza). MSCs were cultured in MSC growth medium (Lonza) containing 10% FBS and 1% GPS until 80% confluence was achieved. MSCs from passage numbers 5 to 8 were used.
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