Journal: Frontiers in Cardiovascular Medicine

Article Title: A refined protocol for the isolation and monoculture of primary mouse renal peritubular endothelial cells

doi: 10.3389/fcvm.2023.1114726

Figure Lengend Snippet: Compositions and components of buffers and solutions for primary MRPEC isolation.

Article Snippet: The following day, cells were gently washed twice with pre-warmed (37°C) 1× PBS –/– to remove cellular debris and non-adherent cells that remained prior to returning them to the incubator with 2 ml of fresh complete MV-EC specific media (EGM-MV2, Lonza).

Techniques: Isolation, Formulation, Activity Assay, Recombinant, Centrifugation, Gradient Centrifugation, Saline, Concentration Assay, Cell Culture

Journal: Histochemistry and Cell Biology

Article Title: Outgrowth, proliferation, viability, angiogenesis and phenotype of primary human endothelial cells in different purchasable endothelial culture media: feed wisely

doi: 10.1007/s00418-019-01815-2

Figure Lengend Snippet: Company, specific media and supplements tested in this study

Article Snippet: Data are given as mean ± SD of 5 (VascuLife; Endopan; MCDB 131) or 6 (ATCC; EGM-MV; PromoCell; Cell Applications) ECFC isolations. a significant vs Cell Application medium

Techniques:

Journal: Histochemistry and Cell Biology

Article Title: Outgrowth, proliferation, viability, angiogenesis and phenotype of primary human endothelial cells in different purchasable endothelial culture media: feed wisely

doi: 10.1007/s00418-019-01815-2

Figure Lengend Snippet: Composition of endothelial cell media tested in this study

Article Snippet: Data are given as mean ± SD of 5 (VascuLife; Endopan; MCDB 131) or 6 (ATCC; EGM-MV; PromoCell; Cell Applications) ECFC isolations. a significant vs Cell Application medium

Techniques:

Journal: Histochemistry and Cell Biology

Article Title: Outgrowth, proliferation, viability, angiogenesis and phenotype of primary human endothelial cells in different purchasable endothelial culture media: feed wisely

doi: 10.1007/s00418-019-01815-2

Figure Lengend Snippet: Effect of different endothelial culture media on colony outgrowth of fpEC. a Colony outgrowth after isolation procedure. Images were taken on day 5 (upper images), 10 (middle images) and 20 (lower images) after isolation using an Olympus CKX41 microscope (× 40 magnification). Scale bar: 200 µm. b Number of days until observation of first colonies after isolation. c Number of colonies at the first day of their observation. Data are given as mean ± SD of five (ATCC; VascuLife; Endopan; MCDB 131) or ten (EGM-MV; PromoCell; Cell Applications) fpEC isolations

Article Snippet: Data are given as mean ± SD of 5 (VascuLife; Endopan; MCDB 131) or 6 (ATCC; EGM-MV; PromoCell; Cell Applications) ECFC isolations. a significant vs Cell Application medium

Techniques: Isolation, Microscopy

Journal: Histochemistry and Cell Biology

Article Title: Outgrowth, proliferation, viability, angiogenesis and phenotype of primary human endothelial cells in different purchasable endothelial culture media: feed wisely

doi: 10.1007/s00418-019-01815-2

Figure Lengend Snippet: Effect of different endothelial culture media on quality of fpEC isolations. Immunocytochemistry for quality control of fpEC isolated in different culture media. Cells were seeded in chamber slides after expansion of first colonies. a fpEC were positive for the endothelial cell markers CD31 and VWF, with highest staining when cells attached very close to each other. b Contaminations with fibroblasts and smooth muscle cells were identified by positive staining for CD90 and SMA, respectively. For negative controls unspecific antibodies of the same isotype were used. Scale bar: 100 µm. c Overview of isolation success, i.e., % isolations without any colony outgrowth, % of outgrowth with contaminations, % of endothelial cell isolations that discontinued to proliferate and % of successful fpEC isolations, as revealed after expansion of the colonies and quality control by immunocytochemical characterisation. Contingency test (Chi square) of successful vs unsuccessful isolations revealed an overall effect of the media with a significance of 0.028. Data are given as % of total numbers of five (ATCC; VascuLife; Endopan; MCDB 131) or ten (EGM-MV; PromoCell; Cell Applications) fpEC isolations

Article Snippet: Data are given as mean ± SD of 5 (VascuLife; Endopan; MCDB 131) or 6 (ATCC; EGM-MV; PromoCell; Cell Applications) ECFC isolations. a significant vs Cell Application medium

Techniques: Immunocytochemistry, Isolation, Staining

Journal: Histochemistry and Cell Biology

Article Title: Outgrowth, proliferation, viability, angiogenesis and phenotype of primary human endothelial cells in different purchasable endothelial culture media: feed wisely

doi: 10.1007/s00418-019-01815-2

Figure Lengend Snippet: Effect of different endothelial culture media on colony outgrowth of ECFC isolated from umbilical cord blood. a Colony outgrowth on day 7 and day 13 after isolation. Scale bar: 200 µm. b Number of days until observation of first colonies after isolation. c Number of colonies per ml cord blood observed on day 7 and 13 after isolation. d Immunocytochemistry for quality control of ECFC isolations. Isolations were pure and without contaminations and thus, always positive for the endothelial cell markers CD31 and VWF and negative for the fibroblast marker CD90 and SMA. For negative controls unspecific antibodies of the same isotype were used. Scale bar: 100 µm. Data are given as mean ± SD of 5 (VascuLife; Endopan; MCDB 131) or 6 (ATCC; EGM-MV; PromoCell; Cell Applications) ECFC isolations. a significant vs Cell Application medium

Article Snippet: Data are given as mean ± SD of 5 (VascuLife; Endopan; MCDB 131) or 6 (ATCC; EGM-MV; PromoCell; Cell Applications) ECFC isolations. a significant vs Cell Application medium

Techniques: Isolation, Immunocytochemistry, Marker