Thromb Haemost 1975; 34(03): 825-839
DOI: 10.1055/s-0038-1654001
Original Article
Schattauer GmbH

Culture of Arterial Endothelial Cells[*]

Characterization and Growth of Bovine Aortic Cells
Francois M Booyse
1   Department of Biochemistry, Rush University, Chicago, Illinois 60612, USA
,
Bonnie J Sedlak
1   Department of Biochemistry, Rush University, Chicago, Illinois 60612, USA
,
Max E Rafelson Jr.
1   Department of Biochemistry, Rush University, Chicago, Illinois 60612, USA
› Author Affiliations
Further Information

Publication History

Received 04 May 1975

Accepted 30 May 1975

Publication Date:
02 July 2018 (online)

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Summary

Arterial endothelial cells were obtained from bovine aortae by mild treatment with collagenase and medium perfusion. These cells were cultured in RPMI-1640 medium containing 15 mM Hepes buffer and 35% fetal calf serum at pH 7.35. Essentially ah (90–95%) the effluent cells were viable and 80% of these cells attached to the substratum within 1 hour. Small patches of attached cells coalesced to form confluent monolayers in 3–5 days. Confluent monolayers of endothelial cells consisted of a homogeneous population of tightly packed, polygonal cells. Selected cultures were serially subcultured (trypsin-EDTA) for 12–14 months (30–35 passages) without any apparent change in morphology or loss of growth characteristics. Primary and three-month old (15 passages) cultures had population doubling times of 32–34 hours and 29–31 hours, respectively. These cells (primary and subcultures) did not require a minimum cell number to become established in culture. Bovine endothelial cells (primary, first, fifth and thirteenth passages) were characterized ultrastructurally by the presence of Weibel-Palade bodies, pinocytotic vesicles and microfilaments and immunologically by the presence of thrombosthenin-like contractile proteins and Factor VIII antigen. The intercellular junctions of post-confluent cultures stained specifically with silver nitrate. From these data, we concluded that identifiable endothelial cells could be obtained from bovine aortae and cultured and maintained for prolonged periods of time.

* An abstract of a portion of this work appeared in Circulation. 1974. 50: (# 4) 111-296.