Isolation and Characterization of Human ECFCs

Human ECFCs were isolated from neonatal cord and peripheral blood and their distinct endothelial phenotypes were verified by flow cytometry as previously described (see Supporting Information Fig. S1) (Hofmann et al., 2009; 2012,; Reinisch and Strunk, 2009 (link, link, link); Reinisch et al., 2009 (link)). HUVECs were obtained from Lonza (Basel, Switzerland). ECFC and HUVECs were grown in endothelial growth medium-2 (EGM-2) (Lonza) containing 2% FBS and 1% penicillin/streptomycin/L-glutamine/heparin (Life Technologies, Carlsbad, CA, USA) and EGM-2 growth factor supplements (composed of bFGF, IGF-2, EGF, VEGF, ascorbic acid, hydrocortisone). Ovarian carcinoma cell lines OVCAR-3 (American Type Culture Collection, Manassas, VA, USA), OVCAR-5 (kindly provided by the Cell Culture Core, Vascular Biology Program, Boston Children's Hospital, Boston, MA, USA) and COV-362 (Sigma Aldrich, St. Louis, MO, USA) were grown in DMEM containing 10% FBS.

Free full text: Click here

Hofmann N.A., Yang J., Trauger S.A., Nakayama H., Huang L., Strunk D., Moses M.A., Klagsbrun M., Bischoff J, & Graier W.F. (2015). The GPR 55 agonist, L-α-lysophosphatidylinositol, mediates ovarian carcinoma cell-induced angiogenesis. British Journal of Pharmacology, 172(16), 4107-4118.

Publication 2015

HUVECs EGM-2 penicillin/streptomycin/L-glutamine/heparin OVCAR-3 OVCAR-5 COV-362

Ascorbic acid Blood Cell culture Cell lines Cord Endothelial Endothelial growth factor Flow cytometry Glutamine Heparin Human Hydrocortisone Igf 2 Neonatal Ovarian carcinoma Penicillin Phenotypes Streptomycin Supplements Vascular Vegf

Corresponding Organization : Boston Children's Hospital

Other organizations : Paracelsus Medical University

Top 5 similar protocols

Variable analysis

independent variables

Isolation of human ECFCs from neonatal cord and peripheral blood

dependent variables

Endothelial phenotypes of isolated human ECFCs verified by flow cytometry

control variables

Culture conditions for ECFCs and HUVECs (endothelial growth medium-2 with 2% FBS, 1% penicillin/streptomycin/L-glutamine/heparin, and EGM-2 growth factor supplements)
Culture conditions for ovarian carcinoma cell lines OVCAR-3, OVCAR-5, and COV-362 (DMEM with 10% FBS)

controls

Positive control: HUVECs, a well-established endothelial cell type
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!