An optimized technique for separation of human decidual leukocytes for cellular and molecular analyses.
OBJECTIVE
Human decidua contains a significant number of leukocytes, phenotypically and functionally different from peripheral blood. In vitro studies of decidual leukocytes require constant improvement of cell isolation technique in order to obtain highly purified viable and functionally active cells.


METHODS
Optimized isolation procedure of decidual leukocytes, based on non-enzymatic cell dispersion, Percoll gradient centrifugation and purification step involving immunomagnetic beads is compared with a similar procedure with Lymphoprep gradient.


RESULTS
The method based on Percoll gradient gave a viable leukocyte fraction with higher amount of alphabetaT- and gammadeltaT cells compared with Lymphoprep gradient. The leukocytes exhibited a well-preserved ultrastructure and surface marker expression and were suitable for molecular analysis. The magnitude and the kinetics of their proliferative response were comparable with peripheral blood lymphocytes.


CONCLUSIONS
The results provide an optimized method for isolation and enrichment of decidual leukocytes, suitable for phenotypic, ultrastructural, molecular and functional analyses.
Nagaeva
Olga
O
Department of Clinical Immunology, Umeå University, Sweden.
Bondestam
Karin
K
Olofsson
Jan
J
Damber
Mats-Göran
MG
Mincheva-Nilsson
Lucia
L
D000911:Antibodies, Monoclonal; D017252:CD3 Complex / Q000235:genetics; D019002:CD56 Antigen / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D003656:Decidua / Q000166:cytology* / Q000276:immunology*; D005260:Female; D005362:Ficoll; D006801:Humans; D018189:Immunomagnetic Separation; D016130:Immunophenotyping; D066298:In Vitro Techniques; D017493:Leukocyte Common Antigens / Q000235:genetics; D007962:Leukocytes / Q000166:cytology* / Q000276:immunology*; D008213:Lymphocyte Activation; D008794:Metrizoic Acid; D016253:Microscopy, Immunoelectron; D011205:Povidone; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000378:metabolism; D012822:Silicon Dioxide; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D019002:CD56 Antigen; D012333:RNA, Messenger; D016693:Receptors, Antigen, T-Cell, alpha-beta; D016692:Receptors, Antigen, T-Cell, gamma-delta; D005362:Ficoll; C016039:Percoll; C038920:lymphoprep; D012822:Silicon Dioxide; D008794:Metrizoic Acid; D017493:Leukocyte Common Antigens; D011205:Povidone