Decreased oxygen supply enhances growth in culture of human mid-trimester amniotic fluid cells

Brackertz, M.; Kubbies, Manfred; Feige, A.; Salk, Darrell

doi:10.1007/bf00292364

Cited by 16 publications

(6 citation statements)

References 22 publications

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“…Traditionally, mammalian cells were cultured under ambient oxygen (20% oxygen). However, it was noted that low oxygen improves the growth of various cell types in culture (Brackertz et al 1983;Bradley et al 1978), and incubator systems became available that allowed the culturing of cells under a physiological oxygen concentration (3% oxygen). The paradox of mouse cell senescence was solved when the group of Judith Campisi demonstrated that mouse fibroblasts do not senesce when cultured in 3% oxygen (Parrinello et al 2003).…”

Section: "Replicative" Senescence Of Mouse Cells Is Caused By Oxidatimentioning

confidence: 99%

Cellular Senescence and Tumor Suppression

2010

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Section: "Replicative" Senescence Of Mouse Cells Is Caused By Oxidatimentioning

confidence: 99%

Cellular Senescence and Tumor Suppression

2010

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“…Many such investigations have used human peripheral blood lymphocytes (PBLs), normally in GO phase of the cell cycle, which were stimulated into cycle by the addition of a mitogen such as phytohaemagglutinin (PHA) (14,(16)(17)(18)(21)(22)(23)(24)(25)(36)(37)(38)(39)(40). Another approach has been to render fibroblasts quiescent by the removal of serum and then to stimulate them by the re-addition of serum (5,6,19,21,35,37,41). Melanoma cells (42), mouse CTLL-cells (17), and rat keratinocytes (34) have also been studied by this method.…”

Section: Data Analysis Synchronised Cellsmentioning

confidence: 99%

“…Apart from direct effects on cells by an anti-BrdUrd antibody labelled with fluoresthe cell cycle, cells become sensitised both to light (8) (5,15,24,36). visualised and, by taking sequential samples, the However, the effect of BrdUrd should always be tested movement of these cells through the cycle can be folusing a conventional DNA histogram obtained by lowed.…”

Section: Cytotoxicity Of Brdurdmentioning

confidence: 99%

Cell cycle analysis of asynchronous cell populations by flow cytometry using bromodeoxyuridine label and Hoechst‐propidium iodide stain

Ormerod

Kubbies²

1992

Cytometry

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Continuous labelling of cells with deoxybromouridine (BrdUrd) followed by staining with a bis-benzimidazole (Hoechst 33258) and a phenanthridinium (propidium iodide or ethidium bromide) allows the cells to be separated by flow cytometry according to the extent of their DNA replication. This BrdUrd-Hoechstl PI method has been used mainly to observe perturbations of the cell cycle in synchronously growing cells. In this paper we demonstrate that, when the method is applied to asynchronously dividing cells, more extensive information can be derived about the effects of cytotoxic a n d other treatments on the kinetics of the cell cycle. The interpretation of the data is explained, the effects of different types of cytotoxic agent are described, and the method is compared briefly to other methods for following cell cycle kinetics. o 1992 Wiley-Liss, Inc.Key terms: Cell cycle kinetics, DNA, counterstaining Knowledge of the proliferative characteristics of cells is important in many areas of biology and oncology and, over the years, many flow cytometric methods have been developed for the study of cell cycle kinetics. The more powerful of these methods have employed 5-bromodeoxyuridine (BrdUrd), which is incorporated into DNA in place of thymidine. Detection of BrdUrd in the cellular DNA enables the fraction of cells in the S phase of their cycle to be enumerated and, using multiparametric flow cytometry, also gives detailed information about the kinetics of the cell cycle.There are three approaches to these measurements. Monoclonal antibodies which react specifically with BrdUrd have been used to provide an analysis of the kinetics of cells pulse-labelled with BrdUrd both in vitro (9,121 and in vivo (1,27,42). Counterstaining with propidium iodide (PI) is used to display cell cycle dependent anti-BrdUrd labelling. The second approach makes use of the observation by Latt (26) that the fluorescence of bis-benzimidazoles (Hoechst 33258 and 33342) bound to DNA is quenched by BrdUrd, the degree of quenching depending on the amount of BrdUrd incorporated (2,211. By staining cells with a combination of Hoechst 33258 and an intercalating dye such as PI or ethidium bromide (EB) whose fluorescence is unaffected by BrdUrd, the resolution of the different compartments of the cell cycle in cells continuously labelled with BrdUrd is greatly improved (3,4,13,20,21,31,32,37). The third technique, which detects BrdUrd in pulse-labelled cells, takes advantage of the decrease of Hoechst and increase of mithramycin fluorescence in the presence of BrdUrd (7). This technique requires more complex data recording due to the subtraction of fluorescence signals in real time.The continuous BrdUrd labelling technique applying the BrdUrd/Hoechst quenching effect has been used to examine the cell cycle kinetics of quiescent cells which have been stimulated into cycle

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“…The inoculum consisted of single cells and cell clumps. Flasks were incubated 3 days at 37°C in a low (10%) oxygen tension humidified 5% CO, incubator [Brackertz et al, 1983;Held and Sonnichsen, 19841. On the fourth day the medium was removed and 5 ml fresh Chang medium was added and cultures were examined daily for growth. When cells reached confluency (generally 7-10 days), the cells were harvested with 0.04% trypsin, passaged into 25 cm2 tissue flasks (passage 1 LP-l]), examined daily, and medium was changed every 3 days.…”

Section: Acquisition Of Chorionic Tissue and Culturementioning

confidence: 99%

Animal models: Prenatal diagnosis of Chediak‐Higashi syndrome in the cat by evaluation of cultured chorionic cells

Kahraman

Prieur

1991

Am. J. Med. Genet.

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The autosomal recessive disease Chediak-Higashi syndrome (CHS) is a progressive and generally fatal disease of humans. The underlying genetic defect in CHS is unknown and prenatal diagnostic methods have not been applied to this disease. The purpose of this study was to determine if CHS chorionic cells expressed a characteristic of CHS--enlarged lysosomes--that would permit the prenatal diagnosis of the disease. Cats with CHS, which have been shown to be homologous with human CHS, were used as the model system in this study. Chorionic tissue samples were obtained from CHS and control cat fetuses and cultures of cells were established. Acid phosphatase was utilized as a marker of lysosomes and cultures of chorionic fibroblasts from CHS and control fetuses were stained histochemically for acid phosphatase. The diameter of the largest lysosomes in 150 cells of each fetus was determined. The mean (+/- SD) diameter (in microns) of the largest lysosomes of normal fetuses was 0.9 +/- 0.13 (range 0.5-7.0 microns), whereas the mean diameter of lysosomes in CHS chorionic cells was 3.9 +/- 0.65 microns (range 0.5-25 microns). These means were significantly different (P less than 0.0001). These data suggest that it should be possible to diagnose human CHS in the first trimester by chorionic villus sampling.

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Decreased oxygen supply enhances growth in culture of human mid-trimester amniotic fluid cells

Cited by 16 publications

References 22 publications

Cellular Senescence and Tumor Suppression

Cellular Senescence and Tumor Suppression

Cell cycle analysis of asynchronous cell populations by flow cytometry using bromodeoxyuridine label and Hoechst‐propidium iodide stain

Animal models: Prenatal diagnosis of Chediak‐Higashi syndrome in the cat by evaluation of cultured chorionic cells

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