The use of antibody specific for bromodeoxyuridine for the immunofluorescent determination of DNA replication in single cells and chromosomes

Gratzner, Howard G.; Leif, Robert C.; Ingram, Drury; Castro, Anthony E.

doi:10.1016/0014-4827(75)90612-6

Cited by 142 publications

(63 citation statements)

References 11 publications

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“…Cell incorporation of BrdU (5-bromo-2′-deoxyuridine) was determined using a cell proliferation kit (GE Healthcare Biosciences, Orsay, France) [57] according to the provider's instructions.…”

Section: Attachment and Proliferation Assaysmentioning

confidence: 99%

Polyelectrolyte Multilayer Films of Controlled Stiffness Modulate Myoblast Cell Differentiation

Ren

Crouzier

Roy

et al. 2008

Adv Funct Materials

244

263

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Beside chemical properties and topographical features, mechanical properties of gels have been recently demonstrated to play an important role in various cellular processes, including cell attachment, proliferation, and differentiation. In this work, we used multilayer films made of poly (L-lysine)/Hyaluronan (PLL/HA) of controlled stiffness to investigate the effects of mechanical properties of thin films on skeletal muscle cells (C2C12 cells) differentiation. Prior to differentiation, cells need to adhere and proliferate in growth medium. Stiff films (E 0 > 320 kPa) promoted formation of focal adhesions and organization of the cytoskeleton as well as an enhanced proliferation, whereas soft films were not favorable for cell anchoring, spreading or proliferation. Then C2C12 cells were switched to a low serum containing medium to induce cell differentiation, which was also greatly dependent on film stiffness. Although myogenin and troponin T expressions were only moderately affected by film stiffness, the morphology of the myotubes exhibited striking stiffness-dependent differences. Soft films allowed differentiation only for few days and the myotubes were very short and thick. Cell clumping followed by aggregates detachment could be observed after ~2 to 4 days. On stiffer films, significantly more elongated and thinner myotubes were observed for up to ~ 2 weeks. Myotube striation was also observed but only for the stiffer films. These results demonstrate that film stiffness modulates deeply adhesion, proliferation and differentiation, each of these processes having its own stiffness requirement.

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Section: Attachment and Proliferation Assaysmentioning

confidence: 99%

Polyelectrolyte Multilayer Films of Controlled Stiffness Modulate Myoblast Cell Differentiation

Ren

Crouzier

Roy

et al. 2008

Adv Funct Materials

244

263

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show abstract

“…It has been suggested that the banding patterns represent the organization of large amounts of repetitive DNA into distinct bands [56]. Antibody to bromodeoxyuridine was used to detect sister chromatid exchange and to follow DNA replication [60].…”

Section: Chromosome Structurementioning

confidence: 99%

Chromatin structure and specificity revealed by immunological techniques

Bustin

1976

FEBS Letters

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“…Here, we removed the infl uence of high-fl ow by 1) normalization of highflow with AVF closure, 2) removal of flow with isolation and organ culture of the artery or 3) with in vitro culture of ECs immediately after the timing considered that ECs began to proliferate. Kinetics of ECs was investigated by bromodeoxyuridine (BrdU) labeling (1,5,(8)(9)(10). We found that ECs once been stimulated by high-fl ow proliferated even after nor-muscular) and ketamine (25 mg/kg, intramuscular) and anesthetized with inhalation anesthesia of sevoflurane (1.5% in O 2 /N 2 O, 2/1 vol/vol).…”

mentioning

confidence: 99%

Normalization of high-flow or removal of flow cannot stop high-flow induced endothelial proliferation

Yamauchi¹,

Takahashi²,

Kobayashi³

et al. 2005

Biomed. Res.

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Endothelial cells (ECs) are activated in response to high-fl ow. Our previous studies using arteriovenous fi stula (AVF) model have demonstrated that high-fl ow in blood vessels induces an early and rapid proliferation of ECs before arterial dilatation. Here, we investigated the proliferation of ECs, which had once been stimulated by high-fl ow loading, in a situation without the infl uence of high-fl ow. First, we induced high-fl ow in the rabbit common carotid artery by using AVF. Then, we removed the infl uence of high-fl ow by normalization of high-fl ow with the closure of AVF or by removal of fl ow itself with tissue isolation and organ culture or with cell culture of ECs, at the timing considered that ECs began to proliferate. Kinetics of ECs was investigated by a laser scanning confocal microscopy, phase-contrast microscopy and light microscopy using bromodeoxyuridine labeling method. We found that ECs, which had once been stimulated by high-flow, transiently proliferated even after normalization of high-fl ow or removal of fl ow. We assume that proliferation of ECs is promised when these cells start to proliferate after high-fl ow loading.Arteries are remodeled in response to a change of blood fl ow or wall shear stress (2, 5). Endothelial cells (ECs) are considered to detect the wall shear stress on the surface of themselves and play an important role in arterial remodeling (4). Our previous studies have demonstrated that high-flow induces the proliferation of arterial ECs before arterial dilatation in dogs, rats and rabbits by using arterio-venous fistula (AVF) model (5-9, 11, 12). This proliferation of ECs is the most distinct and earliest morphological change (5, 9), and is an initial change of the remodeling. However, mechanisms of the proliferation are still uncertain. In our high-flow model performed so far (5, 9), high-fl ow was continuing during the whole experimental period. Therefore, it is not certain whether continuous highflow is necessary during the whole period of the proliferation of ECs or only transient high-fl ow is required at the beginning of the proliferation of ECs. The purpose of this study is to investigate the proliferation of ECs, which have once been stimulated by high-fl ow loading, in a situation without the infl uence of high-fl ow. Here, we removed the infl uence of high-fl ow by 1) normalization of highflow with AVF closure, 2) removal of flow with isolation and organ culture of the artery or 3) with in vitro culture of ECs immediately after the timing considered that ECs began to proliferate. Kinetics of ECs was investigated by bromodeoxyuridine (BrdU) labeling (1,5,(8)(9)(10). We found that ECs once been stimulated by high-fl ow proliferated even after nor-

show abstract

The use of antibody specific for bromodeoxyuridine for the immunofluorescent determination of DNA replication in single cells and chromosomes

Cited by 142 publications

References 11 publications

Polyelectrolyte Multilayer Films of Controlled Stiffness Modulate Myoblast Cell Differentiation

Polyelectrolyte Multilayer Films of Controlled Stiffness Modulate Myoblast Cell Differentiation

Chromatin structure and specificity revealed by immunological techniques

Normalization of high-flow or removal of flow cannot stop high-flow induced endothelial proliferation

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