PMID- 24070690
OWN - NLM
STAT- MEDLINE
DCOM- 20140220
LR  - 20210920
IS  - 1383-5718 (Print)
IS  - 1383-5718 (Linking)
VI  - 758
IP  - 1-2
DP  - 2013 Dec 12
TI  - Comparison of three-colour flow cytometry and slide-based microscopy for the 
      scoring of micronucleated reticulocytes in rat bone-marrow and peripheral blood.
PG  - 12-7
LID - S1383-5718(13)00266-0 [pii]
LID - 10.1016/j.mrgentox.2013.07.010 [doi]
AB  - The aim of this study was to perform the first transferability assessment in 
      China of the micronucleus (MN) scoring method based on three-colour flow 
      cytometry (FCM). This was accomplished for both rat bone-marrow and peripheral 
      blood specimens following exposure to a variety of genotoxic and non-genotoxic 
      chemicals, whereby micronucleus induction was measured both with FCM and with 
      traditional microscopy. In an initial study, rats were treated with vehicle or 
      cyclophosphamide (CP) for 2 consecutive days by oral gavage, and blood and bone 
      marrow were sampled at 24 h after the second treatment. Staining with acridine 
      orange (AO) of methanol-fixed slides was used for microscopical analysis and 2000 
      reticulocytes (RET) or polychromatic erythrocytes (PCE) were scored for MN 
      frequency. The erythrocytes in the remaining bone-marrow cell suspensions were 
      eluted on cellulose columns. The eluted bone marrow as well as the peripheral 
      blood cells was fixed, incubated and analyzed by FCM. In another experiment, the 
      performance of the FCM-MN method was further evaluated with five clastogens 
      (urethane, 5-fluorouracil, mitomycin C, methylmethane sulfonate and 
      6-thioguanine), two aneugens (vincristine sulfate and colchicine) and two 
      non-genotoxic new drugs (compounds A and B), whose results were negative in the 
      routine mouse-micronucleus test (MNT). The MN frequencies in rat peripheral blood 
      induced by the positive chemicals were found to be lower than the frequencies in 
      rat bone-marrow by both scoring methods. However, a high level of agreement for 
      the MN frequencies in both compartments was obtained. Good correspondence between 
      the two analysis methods was also achieved. These data provide support that the 
      three-colour FCM method is more rapid and objective than manual microscopy, while 
      yielding comparable data. It further supports the premise that rat peripheral 
      blood may be an alternative to rat bone marrow in the MNT.
CI  - Copyright (c) 2013 Elsevier B.V. All rights reserved.
FAU - Zhou, Changhui
AU  - Zhou C
AD  - National Shanghai Center for New Drug Safety Evaluation & Research, China State 
      Institute of Pharmaceutical Industry, Shanghai 201203, China.
FAU - Wang, Qingli
AU  - Wang Q
FAU - Wang, Zheng
AU  - Wang Z
FAU - Chang, Yan
AU  - Chang Y
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
DEP - 20130923
PL  - Netherlands
TA  - Mutat Res Genet Toxicol Environ Mutagen
JT  - Mutation research. Genetic toxicology and environmental mutagenesis
JID - 101632149
RN  - 8N3DW7272P (Cyclophosphamide)
SB  - IM
MH  - Animals
MH  - Bone Marrow Cells/*cytology
MH  - Color
MH  - Cyclophosphamide/toxicity
MH  - Flow Cytometry/*methods
MH  - *Micronucleus Tests
MH  - Rats
MH  - Reticulocytes/*ultrastructure
OTO - NOTNLM
OT  - Aneugen
OT  - Clastogen
OT  - Cyclophosphamide
OT  - Genotoxicity
OT  - Micronucleus
OT  - Polychromatic erythrocytes
EDAT- 2013/09/28 06:00
MHDA- 2014/02/22 06:00
CRDT- 2013/09/28 06:00
PHST- 2013/02/22 00:00 [received]
PHST- 2013/05/11 00:00 [revised]
PHST- 2013/07/08 00:00 [accepted]
PHST- 2013/09/28 06:00 [entrez]
PHST- 2013/09/28 06:00 [pubmed]
PHST- 2014/02/22 06:00 [medline]
AID - S1383-5718(13)00266-0 [pii]
AID - 10.1016/j.mrgentox.2013.07.010 [doi]
PST - ppublish
SO  - Mutat Res Genet Toxicol Environ Mutagen. 2013 Dec 12;758(1-2):12-7. doi: 
      10.1016/j.mrgentox.2013.07.010. Epub 2013 Sep 23.