Differentiation of single populations in a bidirectional mixed lymphocyte culture using X and Y chromosome-specific FiSH markers

Objective: To validate a new combination of a viability assay and Fluorescence in Situ Hybridsation (FiSH) techniques using X- or Y-DNA probes to assay the viability of each single population within bidirectional mixed lymphocyte cultures (MLCs). Study design: We determined the absolute viable lymph...

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Bibliographic Details
Published in:Journal of immunological methods 2002-08, Vol.266 (1), p.105-110
Main Authors: Brune, Thomas, Riepe, Felix G, Beier, Kirsten, Exeler, Rita, Louwen, Frank, Garritsen, Henk
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cell Survival
Cells, Cultured
Diverse techniques
Female
Fetal
Fluorescence in Situ Hybridsation (FiSH)
Fundamental and applied biological sciences. Psychology
Genetic Markers
Humans
In Situ Hybridization, Fluorescence - methods
Infant, Newborn
Lymphocyte Activation
Lymphocyte Count
Lymphocyte Culture Test, Mixed - methods
Lymphocytes - cytology
Lymphocytes - metabolism
Lymphocytes - ultrastructure
Male
Maternal
Metaphase
Metaphases
Mixed lymphocyte reaction (MLR)
Molecular and cellular biology
Thymidine - biosynthesis
Tritiated thymidine incorporation assay
X Chromosome
Y Chromosome
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Summary:Objective: To validate a new combination of a viability assay and Fluorescence in Situ Hybridsation (FiSH) techniques using X- or Y-DNA probes to assay the viability of each single population within bidirectional mixed lymphocyte cultures (MLCs). Study design: We determined the absolute viable lymphocyte counts of each population in bidirectional maternal–fetal MLCs as well as in bidirectional MLCs between male and female unrelated adults using a combination of a viability assay and additional sex discrimination by FiSH, and compared the results with the classical tritiated thymidine incorporation assay and with sex discrimination by metaphase number. Results: The results of the total viability assay correlated with those of the tritiated thymidine incorporation assay. The differentiation of each single population by FiSH showed, as in the metaphase assay, that the relationship between viable maternal and neonatal cells was shifted significantly ( p
ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(02)00107-2