Antihemophilic Factor Antigen. LOCALIZATION IN ENDOTHELIAL CELLS BY IMMUNOFLUORESCENT MICROSCOPY

Hoyer, Leon W.; Santos, Rene P. de los; Hoyer, John R.

doi:10.1172/jci107469

Cited by 422 publications

(119 citation statements)

References 35 publications

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“…The addition of either an intact HFN matrix or ECGF to the culture environment did not prevent, but significantly delayed the process of HUV-EC organization. The ceils involved in tube development were endothelial cells and not smooth muscle cell contaminants, as the tubular structures contained the factor VIII:Ag and Weibel-Palade bodies, two markers that are specific for endothelial cells (15,36). Furthermore, ceils derived from tubular structures by either trypsin digestion or tube explant methods grew to form a classic cobblestone endothelial cell monolayer and contained factor VIII:Ag.…”

Section: Discussionmentioning

confidence: 98%

Organizational behavior of human umbilical vein endothelial cells.

Maciag¹,

Kadish²,

Wilkins³

et al. 1982

The Journal of Cell Biology

234

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ABSTRAC-f Culture conditions that favor rapid multiplication of human umbilical vein endothelial cells (HUV-EC) also support long-term serial propagation of the cells. This is routinely achieved when HUV-EC are grown in Medium t99 (M-199) supplemented with fetal bovine serum (FBS) and endothelial cell growth factor (ECGF), on a human fibronectin (HFN) matrix. The HUV-EC can shift from a proliferative to an organized state when the in vitro conditions are changed from those favoring low density proliferation to those supporting high density survival. When ECGF and HFN are omitted, cultures fail to achieve confluence beyond the first or second passage: the preconfluent cultures organize into tubular structures after 4-6 wk. Some tubes become grossly visible and float in the culture medium, remaining tethered to the plastic dish at either end of the tube. On an ultrastructural level, the tubes consist of cells, held together by junctional complexes, arranged so as to form a lumen. The smallest lumens are formed by one cell folding over to form a junction with itself. The cells contain WeibeI-Palade bodies and factor VIII-related antigen. The lumens contain granular, fibrillar and amorphous debris. Predigesting the HFN matrix with trypsin (10 min, 37°C) or plasmin significantly accelerates tube formation. Thrombin and plasminogen activator had no apparent effect. Disruption of the largest tubes with trypsin/EDTA permits the cells to revert to a proliferative state if plated on HFN, in M-199, FBS, and ECGF. These observations indicate that culture conditions that do not favor proliferation permit attainment of a state of nonterminal differentiation (organization) by the endothelial cell. Furthermore, proteolytic modification of the HFN matrix may play an important role in endothelial organization.Normal human diploid cells in culture require serum as a source of hormones and growth factors for cell division (13). The human umbilical vein endothelial cell (HUV-EC) is a fastidious cell with only limited proliferative potential in serum-supplemented medium (11,22). Since serum supplies hormones and growth factors for cell growth in vitro (13,21,38), it can be argued that serum is deficient in the requisite hormone(s) responsible for HUV-EC growth. This approach to HUV-EC growth control has resulted in the identification and characterization of an endothelial cell growth factor (ECGF), partially purified from bovine hypothalamus (20), that stimulates the growth of 22). In the presence of a human fibronectin (HFN) matrix, ECGF permits the growth of HUV-EC at clonal cell densities, reduces the serum requirement for HUV-EC growth, and permits the serial propagation of the HUV-EC (22). One can presently achieve at least 34 cumulative population doublings in an environment composed of a HFN matrix and medium 199 supplemented with ECGF and fetal bovine serum (22).In the absence of HFN and ECGF, HUV-EC do not proliferate beyond 2 or 3 passages. However, under these conditions, the cell population actively organizes into three-...

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Section: Discussionmentioning

confidence: 98%

Organizational behavior of human umbilical vein endothelial cells.

Maciag¹,

Kadish²,

Wilkins³

et al. 1982

The Journal of Cell Biology

234

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“…With the advent of sophisticated immunological techniques, including monoclonal antibodies, it seems likely that (Hoyer et al, 1973) and monoclonal antibodies have recently been raised against this antigen (Sola et al, 1981). It should be possible to conjugate an S-phase-specific chemotherapeutic agent with an endothelial-cell-specific antibody, which could result in a large degree of cell killing in tumour capillaries with very little damage to any normal tissue vessels.…”

mentioning

confidence: 99%

Endothelial cell proliferation as a novel approach to targeting tumour therapy

Denekamp¹

1982

Br J Cancer

227

127

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“…In his studies, vWF antigen was present in endothelial cells in all tissues tested by immunofluorescence but was not present in smooth muscle cells or in fibroblasts (14). In collaboration with him, we studied the expression of vWF antigen by our cultured endothelial cells and were able to demonstrate that the cells in our culture all contained vWF, whereas vWF was absent in all cultured smooth muscle cells and fibroblasts.…”

Section: Figurementioning

confidence: 79%