The tissue suspension was filtered with a 0.22 µm pore-size filter to obtain a monocellular cell suspension. Cells were plated into a 25 cm2 flask coated with bovine type I collagen (BD Biosciences, Milan, Italy) and cultured in endothelial proliferation medium, EndoPM, at 37 °C in 5% CO2 and 5% O2, according to published protocols [5 (link),21 (link),22 (link)], in order to isolate the endothelial cell population (ECs). The media was changed 1–2 times a week and passaged at a split ratio of 1:4 every 14 days [5 (link),9 (link)].
Campanella R., Guarnaccia L., Cordiglieri C., Trombetta E., Caroli M., Carrabba G., La Verde N., Rampini P., Gaudino C., Costa A., Luzzi S., Mantovani G., Locatelli M., Riboni L., Navone S.E, & Marfia G. (2020). Tumor-Educated Platelets and Angiogenesis in Glioblastoma: Another Brick in the Wall for Novel Prognostic and Targetable Biomarkers, Changing the Vision from a Localized Tumor to a Systemic Pathology. Cells, 9(2), 294.
BovineCells Cultured medium Endothelial Endothelial cell Tissue Type i collagen
Corresponding Organization : Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico
Other organizations :
University of Milan, Istituto Nazionale Genetica Molecolare, Ospedale Maggiore, University of Pavia, Policlinico San Matteo Fondazione, Istituti di Ricovero e Cura a Carattere Scientifico, Institute of Biomedical Technologies
Filtration of the tissue suspension using a 0.22 µm pore-size filter
Plating of the cells into a 25 cm^2 flask coated with bovine type I collagen
Culture conditions: 37 °C in 5% CO2 and 5% O2
dependent variables
Isolation of the endothelial cell population (ECs)
control variables
Endothelial proliferation medium, EndoPM
Media change frequency: 1–2 times a week
Passaging at a split ratio of 1:4 every 14 days
Annotations
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