Long-term serial cultivation of arterial and capillary endothelium from adult bovine brain

Goetz, Ingeburg E.; Warren, Jean; Estrada, Carmen; Roberts, Eugene; Krause, Diana N.

doi:10.1007/bf02621355

Cited by 52 publications

(11 citation statements)

References 31 publications

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“…The cells were incubated for 16 h and labeled with [ 3 H]thymidine (5 Ci/ml; NEN Life Science Products) for the last 4 h. The radioactivity incorporated into DNA was determined with a scintillation spectrometer. Bovine capillary endothelial (BCE) cells were isolated from bovine brain cortex as described previously (26). The cells were cultured in RPMI 1640 medium containing 10% FBS supplemented with 15 mM HEPES (pH 7.3) and 2 ng/ml FGF-2 (R&D Systems, Minneapolis, MN) in dishes that had been coated with type IV collagen (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Identification of Chondromodulin I as a Novel Endothelial Cell Growth Inhibitor

Hiraki¹,

Inoue²,

Iyama³

et al. 1997

Journal of Biological Chemistry

170

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Cartilage is unique among tissues of mesenchymal origin in that it is resistant to vascular invasion due to an intrinsic angiogenic inhibitor. During endochondral bone formation, however, calcified cartilage formed in the center of the cartilaginous bone rudiment allows vascular invasion, which initiates the replacement of cartilage by bone. The transition of cartilage from the angioresistant to the angiogenic status thus plays a key role in bone formation. However, the molecular basis of this phenotypic transition of cartilage has been obscure. We report here purification of an endothelial cell growth inhibitor from a guanidine extract of bovine epiphyseal cartilage. The N-terminal amino acid sequence indicated that the inhibitor was identical to chondromodulin I (ChM-I), a cartilage-specific growthmodulating factor. Purified ChM-I inhibited DNA synthesis and proliferation of vascular endothelial cells as well as tube morphogenesis in vitro. Expression of ChM-I cDNA in COS7 cells indicated that mature ChM-I molecules were secreted from the cells after post-translational modifications and cleavage from the transmembrane precursor at the predicted processing signal. Recombinant ChM-I stimulated DNA synthesis and proteoglycan synthesis of cultured growth plate chondrocytes, but inhibited tube morphogenesis of endothelial cells. In situ hybridization and immunohistochemical studies indicated that ChM-I is specifically expressed in the avascular zone of cartilage in developing bone, but not present in calcifying cartilage. These results suggest a regulatory role of ChM-I in vascular invasion during endochondral bone formation.

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Section: Methodsmentioning

confidence: 99%

Identification of Chondromodulin I as a Novel Endothelial Cell Growth Inhibitor

Hiraki¹,

Inoue²,

Iyama³

et al. 1997

Journal of Biological Chemistry

170

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“…Robinson et al (1990) (Goetz et al, 1985). In these studies, the ability of ECs to form capillary-like tubes is dependent on the presence of a specific extracellular matrix or growth factor.…”

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confidence: 98%

Characterization of a γ‐glutamyl transpeptidase positive subpopulation of endothelial cells in a spontaneous tube‐forming clone of rat cerebral resistance‐vessel endothelium

Wang

Grammas

Bault

1993

Journal Cellular Physiology

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A spontaneous tube-forming clone of rat cerebral resistance-vessel endothelium was characterized in long-term serial culture. In this study, a clone, RV-150 ECT, of cerebral resistance vessel endothelial cells in long-term culture has been shown to have a subpopulation of gamma-GTP positive cells that are present in all cultures regardless of confluency status or tube-forming stages. In pre-confluent and confluent cultures, the gamma-GTP positive cells are few in number, stain weakly, and are randomly distributed in the monolayers. In monolayer post-confluent cultures, gamma-GTP positive cells increase in number, stain strongly, and begin to show signs of non-random distributions. In early post-confluent cultures that have become a mixture of monolayer and multilayer cells, there is a further increase in gamma-GTP positive cells which begin to form distinct groupings. In mid post-confluent cultures, the multilayered areas of the culture have begun clustering to form clear multicellular aggregates. The gamma-GTP positive cells at this stage are reduced in number and are predominantly associated with the cell clusters. In late post-confluent cultures, the multicellular clusters develop clear cell cords between/among the clusters. At this stage the gamma-GTP positive cells are associated exclusively with cell clusters. With cord development, the gamma-GTP positive cells are associated with both clusters and cords, and are reduced in number apparently because of selective degeneration of these cells. The results of this study demonstrate that a phenotypically distinct subpopulation of endothelial cells exhibits characteristic features of the blood-brain barrier, namely gamma-GTP. The ability of these cells to express this property in long-term serial culture suggests that this may represent a useful in vitro model to study the growth and differentiation of blood-brain barrier vessels.

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“…A number of morphological and biochemical criteria have been used to identify these cells. These are (1) morphological cobblestone appearance and contact inhibition in culture which has been associated with endothelium in vitro [23]; (2) histological staining for factor VIII antigen which is characteristic of endothelium both in vivo and in vitro and should distinguish these cells from other mesodermally derived cells [14,17,24]. These cells were also negative for GFAP which suggest that they are not of astrocytic origin; (3) EM morphology which demonstrates the presence of intercellular junctional complexes, Weibel-Palade bodies, extensive smooth endoplasmic reticulum and few vesicles.…”

Section: Discussionmentioning

confidence: 99%

Human cerebral endothelium: Isolation and characterization of cells derived from microvessels of non-neoplastic and malignant glial tissue

Costello

Maestro

1990

J Neuro-Oncol

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Human microvessels were isolated and cultured from non-neoplastic cerebral tissue specimens resected for the treatment of seizure disorders and from malignant glial tumors. After 1-2 weeks, cobblestone patterned plaques of cells were isolated and cultured from these microvessels. Cell lines positive for Factor VIII antigen and negative for glial fibrillary acidic protein were designated as endothelium. Endothelium from both tissue sources produced gamma-glutamyl transpeptidase in response to glial cell conditioned media. Tumor derived microvessel endothelium had decreased longevity in culture when compared to normal microvessel endothelium. Tumor derived endothelium also formed less extensive intercellular junctional complexes in vitro. The isolation and characterization of human cerebral microvessel endothelium derived from non-neoplastic tissue and glial tumors may lead to a further understanding of the role of endothelium in tumor growth and vascular permeability alterations.

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Long-term serial cultivation of arterial and capillary endothelium from adult bovine brain

Cited by 52 publications

References 31 publications

Identification of Chondromodulin I as a Novel Endothelial Cell Growth Inhibitor

Identification of Chondromodulin I as a Novel Endothelial Cell Growth Inhibitor

Characterization of a γ‐glutamyl transpeptidase positive subpopulation of endothelial cells in a spontaneous tube‐forming clone of rat cerebral resistance‐vessel endothelium

Human cerebral endothelium: Isolation and characterization of cells derived from microvessels of non-neoplastic and malignant glial tissue

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