PMID- 24548793
OWN - NLM
STAT- MEDLINE
DCOM- 20140516
LR  - 20140520
IS  - 1383-5718 (Print)
IS  - 1383-5718 (Linking)
VI  - 762
DP  - 2014 Mar 1
TI  - Flow cytometric analysis of micronuclei in rat peripheral blood: An 
      interlaboratory reproducibility study.
PG  - 39-42
LID - S1383-5718(14)00034-5 [pii]
LID - 10.1016/j.mrgentox.2014.02.001 [doi]
AB  - In anticipation of proposed OECD guideline changes that may include increasing 
      the number of reticulocytes scored for micronuclei, an inter-laboratory 
      reproducibility study of the rat peripheral blood micronucleus assay was 
      performed using flow cytometry. In this experiment, male Sprague-Dawley (SD) rats 
      were treated with the model clastogen cyclophosphamide (CP: 5, 10 or 15mg/kg) by 
      a single oral administration. As controls, rats were treated with physiological 
      saline (solvent) in the same manner as for the model clastogen. Peripheral blood 
      was collected from each rat 48h after the treatment. The blood samples were 
      prepared at the Public Interest Incorporated Foundation, BioSafety Research 
      Center (BSRC) in duplicate using the rat MicroFlow(PLUS) Kit. After fixation, one 
      replicate set of samples was shipped to Litron Laboratories, and each sample was 
      analyzed by flow cytometry at the two laboratories. In addition, the frequency of 
      micronucleated reticulocytes (MNRETs) was determined at the BSRC by microscopic 
      analysis using supravital acridine orange (AO) staining. The reproducibility of 
      micronucleated reticulocyte frequencies analyzed by microscopy and flow cytometry 
      showed good correlation (r(2)=0.84). The frequencies of micronucleated 
      reticulocytes analyzed by flow cytometry at the two independent laboratories 
      showed good concordance (r(2)=0.97). The data indicate that the flow cytometric 
      micronucleus analysis method is a good alternative to manual microscopic 
      analysis. Flow cytometry allows groups to readily score 5000 or even 20,000 RETs 
      in a matter of minutes compared to manual analysis. This results in increased 
      reliability of the assay by achieving better statistical power.
CI  - Copyright (c) 2014 Elsevier B.V. All rights reserved.
FAU - Kasamoto, Sawako
AU  - Kasamoto S
AD  - Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, 
      Shioshinden, Iwata, Shizuoka 437-1213, Japan. Electronic address: 
      kasamoto-sawako@anpyo.or.jp.
FAU - Mukai, Daisuke
AU  - Mukai D
AD  - Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, 
      Shioshinden, Iwata, Shizuoka 437-1213, Japan.
FAU - Masumori, Shoji
AU  - Masumori S
AD  - Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, 
      Shioshinden, Iwata, Shizuoka 437-1213, Japan.
FAU - Suzuki, Kenichiro
AU  - Suzuki K
AD  - Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, 
      Shioshinden, Iwata, Shizuoka 437-1213, Japan.
FAU - Tanaka, Ryota
AU  - Tanaka R
AD  - Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, 
      Shioshinden, Iwata, Shizuoka 437-1213, Japan.
FAU - Torous, Dorothea K
AU  - Torous DK
AD  - Litron Laboratories, 3500 Winton Place, Rochester, NY 14623, USA.
FAU - Yamate, Jyoji
AU  - Yamate J
AD  - Osaka Prefecture University, 1-58, Rinku Ourai Kita, Izumisano, Osaka 598-8531, 
      Japan.
FAU - Hayashi, Makoto
AU  - Hayashi M
AD  - Public Interest Incorporated Foundation, BioSafety Research Center (BSRC), 582-2, 
      Shioshinden, Iwata, Shizuoka 437-1213, Japan.
LA  - eng
PT  - Journal Article
DEP - 20140215
PL  - Netherlands
TA  - Mutat Res Genet Toxicol Environ Mutagen
JT  - Mutation research. Genetic toxicology and environmental mutagenesis
JID - 101632149
RN  - 0 (Mutagens)
RN  - 8N3DW7272P (Cyclophosphamide)
SB  - IM
MH  - Animals
MH  - Cyclophosphamide/toxicity
MH  - *Flow Cytometry
MH  - Laboratories/*standards
MH  - Male
MH  - Micronuclei, Chromosome-Defective/*chemically induced/statistics & numerical data
MH  - Mutagens/toxicity
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Reproducibility of Results
MH  - Reticulocytes/*drug effects/pathology
MH  - Sensitivity and Specificity
MH  - Toxicity Tests/*standards
OTO - NOTNLM
OT  - Flow cytometry
OT  - Micronucleus assay
OT  - Rat
OT  - Reticulocytes
EDAT- 2014/02/20 06:00
MHDA- 2014/05/17 06:00
CRDT- 2014/02/20 06:00
PHST- 2013/12/11 00:00 [received]
PHST- 2014/02/04 00:00 [revised]
PHST- 2014/02/06 00:00 [accepted]
PHST- 2014/02/20 06:00 [entrez]
PHST- 2014/02/20 06:00 [pubmed]
PHST- 2014/05/17 06:00 [medline]
AID - S1383-5718(14)00034-5 [pii]
AID - 10.1016/j.mrgentox.2014.02.001 [doi]
PST - ppublish
SO  - Mutat Res Genet Toxicol Environ Mutagen. 2014 Mar 1;762:39-42. doi: 
      10.1016/j.mrgentox.2014.02.001. Epub 2014 Feb 15.