A growth history comparison of the human diploid cells WI-38 and IMR-90: Proliferative capacity and cell sizing analysis

Houghton, Bob; Stidworthy, George

doi:10.1007/bf02618249

Cited by 20 publications

(10 citation statements)

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“…Senescence is defined as the loss ofproliferative capacity ofthe cultures and results in the inability of the population to increase in cell number typically after cultures have been passaged 50 or more times. Senescence is associated with a number of gross cellular changes including cell-cycle arrest (3,4), increase in cell size and size heterogeneity (5)(6)(7)(8), and increase in the frequency of cells with chromosomal aberrations, including polyploidy (9)(10)(11)(12). However, molecular data indicate that young and senescent cells do not consistently differ in the pattern of regulation of genes that are sensitive to serum stimulation (13) and several aspects of DNA replication have been reported not to change with culture age (14,15).…”

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Defining cellular senescence in IMR-90 cells: a flow cytometric analysis.

Sherwood

Rush²,

Ellsworth³

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

166

102

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Using multiparameter flow cytometric analysis, we find that senescent cells accumulate in a unique cell-cycle compartment characterized in cell-cycle arrest in G, and a significantly reduced nucleocytoplasmic ratio (genome size/cell mass) relative to cycling cells. With respect to gross cellular phenotype, the quiescent state of senescent cells differs from quiescence induced by density inhibition; the former is associated with a reduction in the nucleocytoplasmic ratio, while the latter is associated with an increase in the nucleocytoplasmic ratio. Senescent cells were present at all passages examined. The frequency of senescent cells was low in earlypassage cultures and increased with passage number. Senescence of populations of IMR-90 cells reflects change in the relative frequency of these cells. The frequency of cells with karyotypic changes increased with the progressive accumulation of out-of-cycle cells.Human diploid fibroblast cells display the phenomenon of cellular senescence and have frequently been used as a model system to study in vitro cellular aging (1, 2). Senescence is defined as the loss ofproliferative capacity ofthe cultures and results in the inability of the population to increase in cell number typically after cultures have been passaged 50 or more times. Senescence is associated with a number of gross cellular changes including cell-cycle arrest (3, 4), increase in cell size and size heterogeneity (5-8), and increase in the frequency of cells with chromosomal aberrations, including polyploidy (9-12). However, molecular data indicate that young and senescent cells do not consistently differ in the pattern of regulation of genes that are sensitive to serum stimulation (13) and several aspects of DNA replication have been reported not to change with culture age (14,15). Thus, while senescent cells may retain normal DNA synthetic capacity, they fail to initiate DNA synthesis and the decline and finally loss of proliferative capacity involves alterations in some process(es) necessary for the initiation of DNA synthesis and/or cell-cycle progression.We have examined the nature of the cell-cycle changes and correlated changes in the frequency of cells with karyotypic abnormalities occurring in IMR-90 cells (16) carried from passage 25 (young cells) to passage 53 (senescent cells). Using multiparameter flow cytometric analysis, we find an age-related increase in the frequency of cells that, based on DNA content, are blocked at the G1/S (diploid and tetraploid) boundary but that become very large (>2 times the population mean) in size relative to cycling cells in the same cell-cycle phase. These large cells fail to label when grown in the presence of bromodeoxyuridine (BrdU) for up to 12 hr while small cells, even in late-passage cultures, do label. Senescent cells thus occupy a unique cell-cycle compartment definable on the basis of altered cell-cycle progression and nucleocytoplasmic ratio. Karyotypic changes associated with cellular aging induced an increase in the frequency of aneupl...

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“…In this figure, the first panel in each row presents the size distribution of the entire cell population (90LS), which as in Fig. 1 (6,21). Senescence of IMR-90 cell populations reflects the progressive change in the relative proportion of these two components of the cell population (22).…”

mentioning

confidence: 99%

Defining cellular senescence in IMR-90 cells: a flow cytometric analysis.

Sherwood

Rush²,

Ellsworth³

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

166

102

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“…In general, since saturation density declines with increasing PDL (Houghton and Stidworthy, 1979;Matuoka and Mitsui, 1981;Macieira-Coelho and Azzarone, 1982;Greenberg et al, 1977; our own data not shown), the results of experiments using older cultures were CM, 6-day conditioned medium containing 10% FBS; 5FM, fresh medium containing 5% FBS; BMFM, fresh medium containing 2M% FBS; 1CM:l S-F, 1 part CM to 1 part serum-free medium; 1CM3 S-F, 1 part CM to 3 parts serum-free medium.…”

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confidence: 95%

Modulation of WI‐38 cell proliferation by elevated levels of CaCl₂

Praeger

Cristofalo

1986

Journal Cellular Physiology

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Elevating the level of extracellular calcium (CaEx2+) increases the saturation density achieved by the normal human diploid cell line, WI-38, but does not change the growth rate. Day 7 cell yields remain unchanged when [CaEx2+] is between 0.5 mM and 3.0 mM, decrease when [CaEx2+] less than 0.5 mM, and increase when [CaEx2+] greater than 3.0 mM. Combining hydrocortisone with additional CaCl2 results in an additive effect on the saturation density relative to that obtained with each treatment separately. The stimulatory effect of elevated [CaCl2] is independent of serum concentration but is lost when WI-38 cells are grown in conditioned medium. Stimulation is recovered when conditioned medium is diluted with serum-free medium. In the case of young cultures grown in conditioned medium, stimulation can also be recovered when higher than usual levels of additional CaCl2 are used (2-3 mM). A glutamine supplementation to the conditioned medium potentiates cell response to elevated [CaCl2]. These results indicate that the loss of an enhanced saturation density when cells are grown in conditioned medium is not due to serum depletion but is more likely the effect of metabolites and/or nutrient depletion. When older or less vigorously growing cultures are grown in conditioned medium, additions of up to 3 mM CaCl2 only lead to inhibition, suggesting an age-related change in proliferative regulation. Elevated levels of CaEx2+ also enhance the proliferative response of quiescent monolayers to serum stimulation. This finding, along with the increase in saturation density of Ca2+-treated cultures, suggests that an elevated level of CaEx2+ affects cell entry into and exit from quiescence brought on by density-dependent inhibition.

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“…The cell life span is defined in the literature as the ability of the epithelial cells to undergo maximum number of divisions in response to injury in order to reconstruct the tissue of origin (Absher et al 1974, Houghton and Stidworthy 1979, Muggleton-Harris and DeSimone 1980 . The average of this maximum number is about 50 .…”

Section: Life Span Of the Stem Cells Tmentioning

confidence: 99%

Transformation of Lung Cells from Inhalation of Radon Daughters in Dwellings: A Preliminary Study

Al-Affan¹,

Haque²

1989

International Journal of Radiation Biology

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There is now world-wide concern for the quantification of lung cancer risk due to indoor radon, but the recent estimates are based on epidemiological studies of miners alone. The present attempt is a preliminary study of the alteration of lung stem cells irradiated by alpha particles emitted by radon daughters. Local energy deposited has been calculated for alpha particles, emitted from radon daughters lining the mucous layer in the respiratory tracts. This calculation has then been followed by dose evaluation and estimation of transformation of lung cells as a function of age. Mean life span of the stem cells was varied between 5 and 45 years to simulate living conditions in different environments. The cumulative fraction of transformed cells after 40 and 70 years has been calculated for radon concentration in the range 23-230 Bq/m3. Increase of the fraction of transformed cells with radon concentration was exponential. It has been assumed that causes other than radiation increase the rate of cell death of mature and stem lung cells, and hence the turnover of stem cells to replace them. It is concluded that the rate of transformation of cells is small for low radon concentration even late in age for non-polluted environments. For radon concentrations of 50 and 100 Bq/m3 the fractions of transformed cells are 0.2 and 6 per cent, respectively for an exposure time of 70 years.

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A growth history comparison of the human diploid cells WI-38 and IMR-90: Proliferative capacity and cell sizing analysis

Cited by 20 publications

References 11 publications

Defining cellular senescence in IMR-90 cells: a flow cytometric analysis.

Defining cellular senescence in IMR-90 cells: a flow cytometric analysis.

Modulation of WI‐38 cell proliferation by elevated levels of CaCl₂

Transformation of Lung Cells from Inhalation of Radon Daughters in Dwellings: A Preliminary Study

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A growth history comparison of the human diploid cells WI-38 and IMR-90: Proliferative capacity and cell sizing analysis

Cited by 20 publications

References 11 publications

Defining cellular senescence in IMR-90 cells: a flow cytometric analysis.

Defining cellular senescence in IMR-90 cells: a flow cytometric analysis.

Modulation of WI‐38 cell proliferation by elevated levels of CaCl2

Transformation of Lung Cells from Inhalation of Radon Daughters in Dwellings: A Preliminary Study

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Modulation of WI‐38 cell proliferation by elevated levels of CaCl₂