Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Optimization of Dendritic cells purification from mouse spleen A recommended method for purification of Dendritic cells from reproductive organs

Authors
Amir Hassan Zarnani 1
Seyyed Mohammad Moazzeni 2
Fazel Shokri 3
Mojdeh Salehnia 4
Ali Ahmad Bayat 5
Mahmood Jeddi-Tehrani 5
1 Immunology Department, Faculty of Medical Science, Tarbiat Modarres University, Tehran, Iran; Nanobiotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran
2 Immunology Department, Faculty of Medical Science, Tarbiat Modarres University, Tehran, Iran
3 Immunology Department, Faculty of Medical Science, Tehran University of Medical Science, Tehran, Iran
4 Anatomy Department, Faculty of Medical Sciences, Tarbiyat Modares University, Tehran, Iran
5 Nanobiotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran
Abstract
Dendritic cells (DC) are the principal antigen-presenting cells (APC) responsible for induction of primary immune responses by T lymphocytes. Although DCs are present in most lymphoid tissues, they occur in very low frequency accounting for 0.5% or less of nucleated cells in peripheral lymphoid organs. In the present study, we report the purification of DCs from mouse spleen with high yield and purity using a three-step purification technique including: collagenase digestion of tissue, selection of low-density cells using Optiprep density gradient medium and plastic adherence. By using techniques outlined above, we obtained 5-7×107 DC/spleen with purity³ of 97%. Such large numbers of purified DCs enables us to further document their different characteristics including morphology, immunophenotype and to evaluation of their role in immune system. Finally, since DCs have been reported to be present in all reproductive organs, we suggest that this protocol be used for isolation and purification of DCs from those organs for further in vitro studies.
Keywords
Dendritic cell
Density gradient centrifugation
Purification
spleen
Journal of Reproduction & Infertility
Volume 4, Issue 1
Winter 2003
Pages 17-29