Chapter 21 Cell-Cycle Analysis Using Continuous Bromodeoxyuridine Labeling and Hoechst 33358—Ethidium Bromide Bivariate Flow Cytometry

Poot, Martin; Hoehn, Holger; Kubbies, Manfred; Grossmann, Angelika; Chen, Yuhchyau; Rabinovitch, Peter S.

doi:10.1016/s0091-679x(08)61726-4

Cited by 38 publications

(46 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Chromosome breakage studies following exposure to various concentrations of mitomycin C were performed according to standard protocols (SchroederKurth et al, 1989). Cell cycle studies employed bivariate BrdU/Hoechst flowcytometry as previously described (Poot et al, 1994;Seyschab et al, 1995). Assignment of patient cells to a given FA complementation group was achieved via complementation analysis using retroviral vectors containing inserts of the full-length cDNAs of FANCA or FANCC (Hanenberg et al, 2002;Lobitz et al, manuscript in preparation).…”

Section: Cell Culture Cell Cycle Studies and Complementation Testingmentioning

confidence: 99%

Reverse mosaicism in Fanconi anemia: natural gene therapy via molecular self-correction

Gross

Hanenberg

Lobitz

et al. 2002

Cytogenet Genome Res

165

123

View full text Add to dashboard Cite

Fanconi anemia (FA) is a genetically and phenotypically heterogenous autosomal recessive disease associated with chromosomal instability and hypersensitivity to DNA crosslinkers. Prognosis is poor due to progressive bone marrow failure and increased risk of neoplasia, but revertant mosaicism may improve survival. Mechanisms of reversion include back mutation, intragenic crossover, gene conversion and compensating deletions/insertions. We describe the types of reversions found in five mosaic FA patients who are compound heterozygotes for single base mutations in FANCA or FANCC. Intragenic crossover could be shown as the mechanism of self-correction in the FANCC patient. Restoration to wildtype via back mutation or gene conversion of either the paternal or maternal allele was observed in the FANCA patients. The sequence environments of these mutations/reversions were indicative of high mutability, and selective advantage of bone marrow precursor cells carrying a completely restored FANCA allele might explain the surprisingly uniform pattern of these reversions. We also describe a first example of in vitro phenotypic reversion via the emergence of a compensating missense mutation 15 amino acids downstream of the constitutional mutation, which explains the reversion to MMC resistance of the respective lymphoblastoid cell line. With one exception, our mosaic patients showed improvement of their hematological status during a three- to six-year observation period, indicating a proliferative advantage of the reverted cell lineages. In patients with Fanconi anemia, genetic instability due to defective caretaker genes sharply increases the risk of neoplasia, but at the same time increases the chance for revertant mosaicism leading to improved bone marrow function.

show abstract

Section: Cell Culture Cell Cycle Studies and Complementation Testingmentioning

confidence: 99%

Reverse mosaicism in Fanconi anemia: natural gene therapy via molecular self-correction

Gross

Hanenberg

Lobitz

et al. 2002

Cytogenet Genome Res

165

123

View full text Add to dashboard Cite

show abstract

“…However, even though chondrocyte proliferation in OA has been reported by others using different probes [1][2][3][4][5], all of them avoid the observation of mitosis as a direct evidence of cell proliferation. Previous studies on proliferation have been based on the incorporation of H 3 thymidine and/or bromodeoxyuridine [6][7][8], which indicates that the cells have passed through S phase of cell cycle during DNA replication, but does not conclusively demonstrate cell division. Likewise, flow cytometry studies could quantify DNA in cell cycle, but was unable to differentiate G 2 phase from M phase [9,10].…”

Section: Introductionmentioning

confidence: 99%

Ontogeny of rat chondrocyte proliferation: studies in embryo, adult and osteoarthritic (OA) cartilage

Gómez-Camarillo

Kourí

2005

Cell Res

View full text Add to dashboard Cite

The aim of this work was to study the ontogeny of chondrocyte cell division using embryo, adult and osteoarthritic (OA) cartilage. We searched for mitosis phases and performed a comparative evaluation of mitotic index, basic fibroblast growth factor b (FGFb), transforming growth factor β1 (TGF-β1) receptors, cyclin dependent kinase (CDK1) and Cyclin-B expression in fetal, neonate, 3, 5, 8 weeks old rats and experimental OA. Our results showed that mitosis phases were observed in all normal cartilage studied, although, we found a decrease in mitotic index in relation to tissue development. No mitosis was detected in OA cartilage. We also found a statistical significant reduction in cell number in OA cartilage, compared with the normal tissue. Furthermore, FGFb and TGF-β1 receptors diminished in relation to tissue development, and were very scarce in experimental OA. Western blot assays showed CDK-1 expression in all cases, including human-OA cartilage. Similar results were observed for Cyclin-B, except for 8 weeks, when it was not expressed. Our results suggest that cell division seems to be scarce, if not absent within the OA cartilage studied. Nevertheless, the existence of factors essential for cell division leaves open the question concerning chondrocyte proliferation in OA cartilage, which is likely to be present in the early stages of the disease.

show abstract

“…Although, generally BrdU infusion as an analog for thymidine nucleotide isused for evaluation of neurogenesis, birth dating, proliferation and cell tracking, but the effects of BrdU on the NSCs behavior have not been fully investigated [15][16][17][18]. For a long-lasting period it has been known that the long term exposure of the NSCs with BrdU promotes conformational changes in DNA structure, affects the balanced nucleotide pool in NSCs and subsequently alters cell cycle progression in which the majority of cells arrest in the G0/G1-phase of the cell cycle [19,23,44]. Also the recent studies have shown that the BrdU alters the DNA structure and stability, raises the risk of sister-chromatid exchange and mutations [18].…”

Section: Discussionmentioning

confidence: 99%

“…Even though, the BrdU has been widely used to label newly generated cells for birth dating, cell fate studies and specially neurogenesis as well as tracking of cells after transplantation but it has some limitations and pitfalls [15][16][17][18]. In the some experimental studies, it has been shown that the administration of the BrdU for a long time induces NSCs senescence, cell death and astroglial differentiation [19][20][21][22][23][24].Therefore, the present study aimed to investigate the potential neurogenesis and regenerative effects of the endogenous neural stem and progenitor cells by means of BrdU injection for labeling in a short time.…”

Section: Introductionmentioning

confidence: 99%

Integration of the Neural Stem and Progenitor Cells into Existing Neuronal Circuitry and Adult Neurogenesis in the Dentate Gyrus of the Hippocampus

Karimipour

Tayefi²,

Shimia³

et al. 2017

j of exper clin neur

View full text Add to dashboard Cite

Chapter 21 Cell-Cycle Analysis Using Continuous Bromodeoxyuridine Labeling and Hoechst 33358—Ethidium Bromide Bivariate Flow Cytometry

Cited by 38 publications

References 29 publications

Reverse mosaicism in Fanconi anemia: natural gene therapy via molecular self-correction

Reverse mosaicism in Fanconi anemia: natural gene therapy via molecular self-correction

Ontogeny of rat chondrocyte proliferation: studies in embryo, adult and osteoarthritic (OA) cartilage

Integration of the Neural Stem and Progenitor Cells into Existing Neuronal Circuitry and Adult Neurogenesis in the Dentate Gyrus of the Hippocampus

Contact Info

Product

Resources

About