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An Optimized Technique for Separation of Human Decidual Leukocytes for Cellular and Molecular Analyses

Authors


Lucia Mincheva-Nilsson, Department of Clinical Immunology, Umeå University, S-90185 Umeå, Sweden. E-mail: lucia.mincheva-nilsson@climi.umu.se

Abstract

PROBLEM: 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.

METHOD OF STUDY: 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 αβT- and γδΤ 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.

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