Proliferative characteristics of clonal endothelial cell strains

Rosen, Eliot M.; Mueller, Stephen N.; Noveral, James P.; Levine, Elliot M.

doi:10.1002/jcp.1041070114

Cited by 92 publications

(38 citation statements)

References 47 publications

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“…These observations suggest that the HUV endothelial cells were either becoming broader or actually increasing in volume as a function of in vitro age. A decrease in confluent cell density with increasing age has also been demonstrated in bovine fetal aortic endothelial cells for which a measured increase in cell attachment area corresponds to larger cell volumes in older populations (26) . The increasing frequency of extremely large HUV endothelial cells at later passages probably accounts for much of the decrease in confluent cell density in our older cultures .…”

Section: Huv Endothelial-cell Growth In Various Seramentioning

confidence: 68%

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Serial propagation of human endothelial cells in vitro.

Maciag¹,

Hoover²,

Stemerman³

et al. 1981

The Journal of Cell Biology

330

149

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Human umbilical vein (HUV) endothelial cells were grown for 15 to 21 passages at a split ratio of 1:5 (at least 27 population doublings) on a human fibronectin (HFN) matrix in Medium 199 supplemented with fetal bovine serum (FBS) and endothelial-cell growth factor (ECGF). This system also permitted the growth of HUV endothelial cells at cell densities as low as 1.25 cells/cm2. In addition to delaying the premature senescence of HUV endothelial cells, ECGF also reduced the serum requirement for low-density HUV endothelial-cell growth; 2.5% serum and ECGF yields half-maximum growth as compared to high serum controls. Significant HUV endothelial-cell growth was also observed in medium supplemented with either ovine hypophysectomized (HYPOX) serum, plasma-derived serum (PDS), or HYPOX-PDS in the presence of ECGF, suggesting that neither the pituitary nor the platelet contributes to HUV endothelial-cell growth.

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Section: Huv Endothelial-cell Growth In Various Seramentioning

confidence: 68%

“…Bovine (1, 3,5,11,22,26) and porcine (14,31) aortic endothelial cells are widely studied due to the relative ease with which they can be propagated in long-term culture . Although endothelial cells from human pulmonary arteries and veins have been grown in culture (19,28), umbilical veins are the most readily available source of human large-vessel endothelium.…”

mentioning

confidence: 99%

Serial propagation of human endothelial cells in vitro.

Maciag¹,

Hoover²,

Stemerman³

et al. 1981

The Journal of Cell Biology

330

149

View full text Add to dashboard Cite

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“…When the harvested cell number did not increase compared to the plated cell number, culture was considered senescent. The PD at each subculture was calculated with the formula PD ϭ log 2 [(number of cells harvested)/(number of cells seeded ϫ attachment efficiency)] and summed to obtain the cumulative PD (Rosen et al, 1981). In this study, attachment efficiency was above 95%.…”

Section: Methodsmentioning

confidence: 99%

Possible involvement of p21 but not of p16 or p53 in keratinocyte senescence

et al. 1999

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It has been reported that p21, p53, and p16 affect the cell cycle and cell senescence. However, their roles in keratinocyte senescence are not clear. We established primary keratinocyte strains from 15 donors and maintained them until replicative senescence; their population doublings ranged from 5.7-45.2. These strains were classified based on their population doublings as short (5.7-10.4), intermediate (13.9-17.4), and long (21.5-45.2). To investigate the roles of p21, p53, and p16 in the cellular senescence of the cultured keratinocytes, we quantitatively analyzed p21, p53, and p16 levels of keratinocyte strains with different life spans by Western blot with Fluorol mager. p21 levels increased in the senescent phase but not in the nonsenescent phase in all of the short, intermediate, and long life-span strains. Northern blot analysis also revealed induction of p21 mRNA was similar to that of p21 protein levels. There were no apparent differences in p53 levels between senescent and nonsenescent cells. The short life-span strains exhibited a significant increase in p16 levels in the senescent phase (eighth or tenth passage). However, in two long life-span strains, p16 levels were increased in the nonsenescent phase (eighth passage) but then declined as the cells reached senescence (twenty-seventh passage). Therefore, induction of p16 appeared not to be associated with senescence in long life-span strains. In conclusion, p21 but not p16 or p53 may play roles in keratinocyte senescence.

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“…We Rosen et al, 1981) and other investigators (Duthu and Smith, 1980;Eskin et al, 1978;Feneslau and Mello, 1976;Gospodarowicz et al, 1981;McAuslan et al, 1979;Ryan et al, 1978;Schwartz, 1978) have provided repeated evidence for the finite in vitro proliferative lifespan of bovine vascular endothelial cells. Spontaneous transformation of cultures into lines with indefinite lifespans has been reported in a few instances for some bovine cell types (Courtois et al, 1978;Madin and Darby, 1958;McClurkin et al, 1974) but has not been reported in bovine endothelial cells and must be presumed to occur only at a low frequency, if at all, in this cell type.…”

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confidence: 89%

Bovine endothelial cells transformed in vitro by benzo(a)pyrene

Grinspan

Mueller

Levine

1983

Journal Cellular Physiology

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A cloned strain of bovine vascular endothelial cells with a finite in vitro lifespan was treated with benzo(a)pyrene (BP) after approximately 75% of its lifespan was completed. Untreated cultures of this strain senesced upon serial subcultivation and contained large, nondividing cells. In three out of seven trials, BP treatment produced transformed lines with altered phenotypic characteristics. The transformed cells appeared in the cultures concomitant with the senescence of the parent cells. All transformed cell lines examined exhibited indefinite lifespans and altered karyotypes. Two of the lines retained most of the characteristics of normal endothelial cells, except that one became aneuploid and the other polyploid. Neither of these lines formed tumors when inoculated into nude mice. The remaining two lines retained mostly diploid karyotypes, but a high percentage of cells contained Robertsonian translocations. In one line cell volume was markedly reduced. In addition, these lines grew in multilayers, were anchorage independent, and proliferated in medium containing 0.5% serum. When 10(7) cells of these lines were injected into nude mice, tumors appeared within 1 week and were identified as malignant hemangioendotheliomas of bovine origin.

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Proliferative characteristics of clonal endothelial cell strains

Cited by 92 publications

References 47 publications

Serial propagation of human endothelial cells in vitro.

Serial propagation of human endothelial cells in vitro.

Possible involvement of p21 but not of p16 or p53 in keratinocyte senescence

Bovine endothelial cells transformed in vitro by benzo(a)pyrene

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