Table of Contents
Bone Marrow Research
Volume 2018, Article ID 3495086, 9 pages
https://doi.org/10.1155/2018/3495086
Research Article

Optimization of Ex Vivo Murine Bone Marrow Derived Immature Dendritic Cells: A Comparative Analysis of Flask Culture Method and Mouse CD11c Positive Selection Kit Method

1Department of Zoonosis, Haffkine Institute for Training, Research and Testing, Parel, Mumbai, India
2Department of Biochemistry and Virology, National Institute for Research in Reproductive Health (NIRRH), Parel, Mumbai, India
3Department of Virology and Immunology, Haffkine Institute for Training, Research and Testing, Parel, Mumbai, India
4Department of Microbiology, Grant Medical College and Sir JJ Group of Hospital, Byculla, Mumbai, India

Correspondence should be addressed to Rahul Ashok Gosavi; moc.liamg@luhar.ivasog and Vainav Patel; moc.liamg@pvaniav

Received 29 August 2017; Revised 24 November 2017; Accepted 7 December 2017; Published 22 February 2018

Academic Editor: Paolo De Fabritiis

Copyright © 2018 Rahul Ashok Gosavi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

12–14 days of culturing of bone marrow (BM) cells containing various growth factors is widely used method for generating dendritic cells (DCs) from suspended cell population. Here we compared flask culture method and commercially available CD11c Positive Selection kit method. Immature BMDCs’ purity of adherent as well as suspended cell population was generated in the decreasing concentration of recombinant-murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF) in nontreated tissue culture flasks. The expression of CD11c, MHCII, CD40, and CD86 was measured by flow cytometry. We found significant difference between the two methods in the adherent cells population but no significant difference was observed between the suspended cell populations with respect to CD11c+ count. However, CD11c+ was significantly higher in both adhered and suspended cell population by culture method but kit method gave more CD11c+ from suspended cells population only. On the other hand, using both methods, immature DC expressed moderate level of MHC class II molecules as well as low levels of CD40 and CD86. Our findings suggest that widely used culture method gives the best results in terms of yield, viability, and purity of BMDCs from both adherent and suspended cell population whereas kit method works well for suspended cell population.