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Improvements of the Bromodeoxyuridine-DNA Flow Cytometry Method for the Study of Cell Proliferation

Johansson, Maria C LU (1999)
Abstract
Potential doubling time (Tpot), DNA synthesis time (TS), and labelling index (LI) are fundamental growth kinetics parameters in clinical and experimental cancer research, which may be of further practical importance regarding prognosis and treatment prediction of cancer. They can be measured by bromodeoxyuridine(BrdUrd)/flow cytometry (FCM) methods, where BrdUrd, an analogue of thymidine, is incorporated into DNA and quantified simultaneously with the DNA content. However, this method requires improvements. Since in many applications only a single sample is available, the method must be reproducible, accurate, and independent of the time of sample collection in relation to BrdUrd pulse-labelling. With the modifications we describe, growth... (More)
Potential doubling time (Tpot), DNA synthesis time (TS), and labelling index (LI) are fundamental growth kinetics parameters in clinical and experimental cancer research, which may be of further practical importance regarding prognosis and treatment prediction of cancer. They can be measured by bromodeoxyuridine(BrdUrd)/flow cytometry (FCM) methods, where BrdUrd, an analogue of thymidine, is incorporated into DNA and quantified simultaneously with the DNA content. However, this method requires improvements. Since in many applications only a single sample is available, the method must be reproducible, accurate, and independent of the time of sample collection in relation to BrdUrd pulse-labelling. With the modifications we describe, growth kinetic data could be obtained in response to the demands, both in vitro and in vivo and they were in agreement with those obtained with the [3H]thymidine/autoradiography method. Thus, the BrdUrd/FCM method can replace traditional [3H]thymidine-based methods. The modifications included new mathematic formulas for the calculation of LI and TS. They were compared with various other formulas, in several cell lines and experimental tumours. Our formulas did show sampling time independence in several cell lines studied. The labelling time should be kept as short as possible. The proposed TS formula is used preferable in more slowly growing cell populations. Tpot values based on our formulas did not depend on sampling time. In conclusion, with our modified BrdUrd/FCM method for growth kinetic studies, experimentally and/or clinically, it is possible to obtain reproducible and sampling time independent data from only one sampling, an advantage of great importance when clinical applications are concerned. (Less)
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author
opponent
  • Ph D Stål, Olle, Linköping University, Sweden
organization
publishing date
type
Thesis
publication status
published
subject
keywords
head and neck cancer, [3H]thymidine, energy transfer, cell cycle, xenografted tumours, cultured cells, potential doubling time, labelling index, DNA synthesis time, cell proliferation, growth kinetics, Bromodeoxyuridine, DNA flow cytometry, cancer, onkologi, Cytologi, cancerology, oncology, Cytology
pages
171 pages
publisher
The Jubileum Institute, Department of Oncology, Lund University Hospital, S-221 85 LUND, Sweden
defense location
The lecture hall, the Oncology department, Lund University Hospital
defense date
1999-02-04 09:00
external identifiers
  • other:ISRN: LUMEDW/MEOK-1024-SE
ISBN
91-628-3348-0
language
English
LU publication?
yes
id
7e2d84b1-9e96-42bb-b3fe-0b198ef43032 (old id 39276)
date added to LUP
2007-06-21 10:39:29
date last changed
2016-09-19 08:45:07
@phdthesis{7e2d84b1-9e96-42bb-b3fe-0b198ef43032,
  abstract     = {Potential doubling time (Tpot), DNA synthesis time (TS), and labelling index (LI) are fundamental growth kinetics parameters in clinical and experimental cancer research, which may be of further practical importance regarding prognosis and treatment prediction of cancer. They can be measured by bromodeoxyuridine(BrdUrd)/flow cytometry (FCM) methods, where BrdUrd, an analogue of thymidine, is incorporated into DNA and quantified simultaneously with the DNA content. However, this method requires improvements. Since in many applications only a single sample is available, the method must be reproducible, accurate, and independent of the time of sample collection in relation to BrdUrd pulse-labelling. With the modifications we describe, growth kinetic data could be obtained in response to the demands, both in vitro and in vivo and they were in agreement with those obtained with the [3H]thymidine/autoradiography method. Thus, the BrdUrd/FCM method can replace traditional [3H]thymidine-based methods. The modifications included new mathematic formulas for the calculation of LI and TS. They were compared with various other formulas, in several cell lines and experimental tumours. Our formulas did show sampling time independence in several cell lines studied. The labelling time should be kept as short as possible. The proposed TS formula is used preferable in more slowly growing cell populations. Tpot values based on our formulas did not depend on sampling time. In conclusion, with our modified BrdUrd/FCM method for growth kinetic studies, experimentally and/or clinically, it is possible to obtain reproducible and sampling time independent data from only one sampling, an advantage of great importance when clinical applications are concerned.},
  author       = {Johansson, Maria C},
  isbn         = {91-628-3348-0},
  keyword      = {head and neck cancer,[3H]thymidine,energy transfer,cell cycle,xenografted tumours,cultured cells,potential doubling time,labelling index,DNA synthesis time,cell proliferation,growth kinetics,Bromodeoxyuridine,DNA flow cytometry,cancer,onkologi,Cytologi,cancerology,oncology,Cytology},
  language     = {eng},
  pages        = {171},
  publisher    = {The Jubileum Institute, Department of Oncology, Lund University Hospital, S-221 85 LUND, Sweden},
  school       = {Lund University},
  title        = {Improvements of the Bromodeoxyuridine-DNA Flow Cytometry Method for the Study of Cell Proliferation},
  year         = {1999},
}