A method for preparing chromosomes from peripheral blood of the mouse

Barren, P.R.; Morris, Suzanne M.; Schol, Henry M.; Bishop, Jack B.

doi:10.1016/0165-7992(82)90138-5

Cited by 5 publications

(4 citation statements)

References 2 publications

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“…In the few previous reports by other investigators culturing mouse IL PBLs for cytogenetic endpoints [Barren et al, 1982;Goon and Conner, 19841 2-ME is used as a growth supplement with some success. Goon and Conner [1984] state that 2-ME is required to obtain successful growth of mouse IL PBLs, although MI data were not presented.…”

Section: Resultsmentioning

confidence: 99%

A modified mouse peripheral blood lymphocyte culture system for cytogenetic analysis

Erexson

Kligerman

1987

Environ and Mol Mutagen

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A detailed methodology is presented for culturing mouse peripheral blood lymphocytes isolated on density gradients and stimulated to divide using either phytohemagglutinin, concanavalin A, or lipopolysaccharide. The techniques described yield more than sufficient numbers of mitotic cells for analyzing sister chromatid exchange, chromosome aberrations, and micronuclei following in vitro or in vivo exposure to chemicals or radiation.

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Section: Resultsmentioning

confidence: 99%

A modified mouse peripheral blood lymphocyte culture system for cytogenetic analysis

Erexson

Kligerman

1987

Environ and Mol Mutagen

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“…For human PBLs, it is generally accepted that 48 hr-culture gives a maximal number of first in vitro metaphase cells [15]. Comparable knowledge was lacking for the mouse PBL culture system, and culture periods ranged from 36 to 72 hr in previous studies [1,3,4,12,16]. Under the culture conditions of the present study, 42 hr-cultures produced little yields of second in vitro G 2 /M cells (Table 2), and longer cultures carried the risk of counting them.…”

Section: Cultures For Conventional Cytogenetic Analysismentioning

confidence: 99%

“…However, many of these culture techniques are laborious and complex and some appear to have enjoyed only limited success. Among these studies, a few studies had somewhat better success obtaining metaphase chromosomes from mouse PBLs using whole blood culture systems [1,2], which were later modified by isolating lymphocytes using Ficoll-Hypaque density gradient [3,4].…”

Section: Introductionmentioning

confidence: 99%

An Improved Culture System of Mouse Peripheral Blood Lymphocytes for Analysis of Radiation-Induced Chromosome Aberrations

Kanda¹,

Shang²,

Tsuji³

et al. 2004

Bioscience Reports

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There is an incentive to develop a culture system of mouse peripheral blood lymphocytes (PBLs) to serve as models for studying genotoxic effects in humans exposed to mutagens, including ionizing radiation. However, many past approaches have been laborious, complex and only partly reproducible. In the present study, we established an improved culture system of mouse PBLs by removing blood and/or plasma, which was found to inhibit in vitro mitotic stimulation or proceeding cell cycles of lymphocytes. We compared the reactions of isolated PBLs to mitogens between the classical method and the present improved one. Then, we applied this method to the cytogenetic analysis using chemically induced premature chromosome condensation (PCC) as well as the conventional analysis, and demonstrated that the frequency of excess fragments observed in PCC cells might be useful to quantify the radiation-induced damages on chromosomes.

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“…For those animals which lacked meiotic metaphase-I cells, peripheral blood was obtained by orbital sinus puncture and cultured according to the methods of Barren et al [1982]. Mitotic metaphase cells were trypsin-banded by a modified Seabright [ 19711 method and karyotyped according to Nesbitt and Franke [ 19731.…”

Section: Cytogenetic Evaluationmentioning

confidence: 99%

Detection of TEM‐induced reciprocal translocations in F₁ sons of CD‐1 male mice: Comparison of sequential fertility evaluation and cytogenetic analysis

Morris¹,

Kodell

Domon

et al. 1988

Environ and Mol Mutagen

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To determine the positive and negative classification error rates associated with the HTA in our laboratory, F1 sons of TEM-exposed CD-1 male mice were evaluated by the sequential fertility method with subsequent cytogenetic analysis. Males who sired three litters of size 10 or less when mated to primiparous females from either the B6C3F1 or the BCF1 strain were classified as partial steriles. When meiotic chromosome analyses revealed the presence of at least two cells containing multivalent figures, males were classified as translocation heterozygotes. When the fertility evaluation and the cytogenetic analysis were compared, normal fertility was observed on 5 of 83 (6.02%) translocation-bearing F1 males mated to B6C3F1 tester females and on 3 of 83 (3.61%) F1 males mated to BCF1 tester females. Thus, the false-negative error rates were 6.02% and 3.61% with these two tester strains. Multivalent figures were not observed in the meiotic chromosomes of 410 F1 males. Of these, 12 (2.93%) had reduced fertility when mated to the B6C3F1 tester strain as did 7 (1.71%) mated to the BCF1 strain. Thus, the false-positive error rates with these two tester strains were 2.93% for the B6C3F1 strain and 1.71% for the BCF1 strain. Our results indicate that non-zero error rates, both false-positive and false-negative, are associated with the sequential mating method HTA. In addition, the magnitude of these error rates was influenced not only by the tester female strain but also by the genotype of the F1 male.

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A method for preparing chromosomes from peripheral blood of the mouse

Cited by 5 publications

References 2 publications

A modified mouse peripheral blood lymphocyte culture system for cytogenetic analysis

A modified mouse peripheral blood lymphocyte culture system for cytogenetic analysis

An Improved Culture System of Mouse Peripheral Blood Lymphocytes for Analysis of Radiation-Induced Chromosome Aberrations

Detection of TEM‐induced reciprocal translocations in F₁ sons of CD‐1 male mice: Comparison of sequential fertility evaluation and cytogenetic analysis

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A method for preparing chromosomes from peripheral blood of the mouse

Cited by 5 publications

References 2 publications

A modified mouse peripheral blood lymphocyte culture system for cytogenetic analysis

A modified mouse peripheral blood lymphocyte culture system for cytogenetic analysis

An Improved Culture System of Mouse Peripheral Blood Lymphocytes for Analysis of Radiation-Induced Chromosome Aberrations

Detection of TEM‐induced reciprocal translocations in F1 sons of CD‐1 male mice: Comparison of sequential fertility evaluation and cytogenetic analysis

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Detection of TEM‐induced reciprocal translocations in F₁ sons of CD‐1 male mice: Comparison of sequential fertility evaluation and cytogenetic analysis