Isolation of Murine Aortic Endothelial Cells in Culture and the Effects of Sex Steroids on Their Growth

Abstract

The lack of commercially available primary murine endothelial cells prompted us to isolate and cultivate this cell type. We report here the effect of sex steroids on the in vitro growth of murine aortic endothelial cells. Murine aortic endothelial cells were isolated by a combination of explant outgrowth from aortic rings and enzymatic digestion. The endothelial nature of the cells was verified by uptake of acylated low-density lipoprotein and positive staining for CD-31. Murine aortic endothelial cell growth is stimulated by physiological concentrations of estrogen. Progesterone, when given simultaneously with estrogen, inhibited the stimulatory growth effect of estrogen. Murine aortic endothelial cells grown in vitro continue to express messenger ribonucleic acid for proteins related to endothelial growth. These include vascular endothelial growth factor, its receptors Flt-1 and Flk-1, and the angiogenesis-associated transcription factor, Ets-1.

Cytocompatibility of electrospun nanofiber tubular scaffolds for small diameter tissue engineering blood vessels

“…Besides, a modified method for isolating SDRAECs was established by us in the way that more highly purified SDRAECs could be harvested in less time than traditional methods including enzyme digestion and tissue attached method [38,39]. The adventitia and media of rat aortas were cleared in order to reduce contamination of fibroblasts and smooth muscle cells.…”

Fibrinogen induces alterations of endothelial cell tight junction proteins

“…Rat heart microvascular endothelial cells (RHMEC) were purchased from Vec Technologies (Rensselaer, NY). The endothelial nature of the cells was verified by uptake of acylated low‐density lipoprotein and positive staining for CD‐31 (Lincoln et al, 2003). The RHMECs were grown in MCDB‐131 Complete medium (Vec Technologies, NY) at 37°C with 5% CO 2 in air in a humidified environment and were used at the 5th or 6th passage for the experiments.…”

Protection of vascular cells from oxidative stress by proteasome inhibition depends on Nrf2

“…Human aortic endothelial cells (HAEC) were obtained from Promocell and cultivated in the presence of a specific endothelial cell growth medium (EC Medium, Promocell) containing 0.4% ECGS/ H, 2% FCS, 0.1 ng/mL epidermal growth factor, 1 mg/mL hydrocortisone, 1 ng/mL basic fibroblast growth factor (Promocell), and 1% penicillin/streptomycin. Murine aortic endothelial cells (MAEC) were prepared from adult mice as described previously 37 and cultivated in a 1:1 mixture of MCDB131 and DMEM (Invitrogen) containing 5% FCS (Biochrom). Vascular smooth muscles cells (mVSMC) were prepared from adult mice as described previously 38 and cultivated in DMEM containing 10% FCS and 1% penicillin/streptomycin.…”