Establishment and Characterization of an Angiosarcoma-Derived Cell Line, AS-M

Abstract

A novel human endothelial cell line, AS-M, has been established from a cutaneous angiosarcoma on the scalp. The cells expressing platelet endothelial cell adhesion molecule-1 (CD31) were isolated using magnetic beads and subsequently cultured for a year. To date, the cells have undergone more than 100 population doublings (PDs). The AS-M cells manifested endothelial characteristics, such as active uptake of acetylated low-density lipoprotein labeled with 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil-Ac-LDL), capacity to bind the Ulex europeaus agglutin-I (UEA-I), and expression of von Willebrand factor (vWF) and CD31. The single cell-derived clone, AS-M.5, showed a constitutive expression of CD31, vWF, angiotensin-converting enzyme (ACE), endoglin (CD105), and the endothelial cell receptor tyrosine kinases KDR and Tie-1. Similarly to freshly isolated endothelial cells, the AS-M.5 responded to induction by bacterial lipopolysaccharide (LPS) by increased transcription of cell adhesion molecules and cytokines. The AS-M.5 cultures required endothelial growth supplements for optimal growth and long-term propagation in vitro. However, in contrast to normal endothelial cells, p53 gene products were detected in nuclei of AS-M.5 cells. Cytogenetic analyses consistently revealed a hypodiploid karyotype with complete loss of one homologue of several chromosomes and a homogeneous pattern of distinct karyotypic changes. Although the AS-M.5 presented characteristics suggestive of tumor cells, they did not develop into tumors when inoculated subcutaneously into nude mice. The cell line AS-M.5 could be a useful model system to study endothelial pathobiology in vitro.

Generation of a murine hepatic angiosarcoma cell line and reproducible mouse tumor model

“…Two human AS cell lines (ISO-HAS and AS-M) were established from human scalp AS. 27,28 The AS cell line ISOS-1 was established from human scalp AS transplanted into immunodeficient mice. 29 Furthermore, a total of seven canine AS cell lines were established from three different xenograft canine tumors.…”

Vascular-targeted agents for the treatment of angiosarcoma

“…Protocols 3 and 4 detailed below were based on methods described by Krump-Konvalinkova et al [154] and Masuzawa et al [156], yielding ASM and ISO-HAS respectively.…”

“…ASM and ISO-HAS have been identified as malignant endothelial cell lines [154,156,158]. Characterisation studies were performed to confirm that ASM and ISO-HAS cells …”

Vascular tumors have increased p70 S6-kinase activation and are inhibited by topical rapamycin

“…Human dermal microvascular ECs (HDMECs) were purified as described previously. 32 Human ASM.5 angiosarcoma cells were a gift from Krump-Konvalinkova et al 33 This cell line was authenticated by DNA fingerprinting in our laboratory. Mouse EOMA and bEND.3 cells were obtained from ATCC.…”

“…Primary infantile hemangioma ECs were purified from fresh hemangioma tissues; the cell purity was determined by the uptake of DiI-acetylated LDL, and by staining for the endothelial markers CD31 and VE cadherin (Supplementary Figure 2). ASM.5 cells were derived from a spontaneous human angiosarcoma 33 and EOMA cells were derived from a spontaneous mouse hemangioendothelioma. 34 bEND.3 cells are mouse brain EC transformed with the polyoma virus middle T antigen.…”