Isolation of Pulmonary Endothelial Cells

Endothelial cells were isolated from the lungs of 4- to 6-week-old mice; although they were younger than mice used in our other experiments, we found that juvenile lungs produced higher-quality cell preparations for functional studies. As previously described, CD146⁺ pulmonary endothelial cells (PECs) were isolated with immunomagnetic microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) and cultured in MV2 medium (Promocell), supplemented with 10% fetal calf serum, 100 units/mL penicillin, and 100 mg/mL streptomycin (9 (link)). Cells were studied at passage 2 to 3.

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Sengupta A., Patel P.A., Yuldasheva N.Y., Mughal R.S., Galloway S., Viswambharan H., Walker A.M., Aziz A., Smith J., Ali N., Mercer B.N., Imrie H., Sukumar P., Wheatcroft S.B., Kearney M.T, & Cubbon R.M. (2018). Endothelial Insulin Receptor Restoration Rescues Vascular Function in Male Insulin Receptor Haploinsufficient Mice. Endocrinology, 159(8), 2917-2925.

Publication 2018

immunomagnetic microbeads MV2 medium

Cells Cultured medium Endothelial cells Fetal calf serum Mice Microbeads Penicillin Pulmonary Streptomycin Younger

Corresponding Organization : University of Leeds

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Variable analysis

independent variables

Age of mice (4- to 6-week-old)

dependent variables

Quality of cell preparations for functional studies

control variables

Passage of cells (2 to 3)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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