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A new protocol for the propagation of dendritic cells from rat bone marrow using recombinant GM-CSF, and their quantification using the mAb OX-62

Chen-Woan, M and Delaney, CP and Fournier, V and Wakizaka, Y and Murase, N and Fung, J and Starzl, TE and Demetris, AJ (1995) A new protocol for the propagation of dendritic cells from rat bone marrow using recombinant GM-CSF, and their quantification using the mAb OX-62. Journal of Immunological Methods, 178 (2). 157 - 171. ISSN 0022-1759

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Abstract

Bone marrow (BM)-derived dendritic cells (DC) are the most potent known antigen (Ag) presenting cell in vivo and in vitro. Detailed analysis of their properties and mechanisms of action requires an ability to produce large numbers of DC. Although DC have been isolated from several rat tissues, including BM, the yield is uniformly low. We describe a simple method for the propagation of large numbers of DC from rat BM and document cell yield with the rat DC marker, OX-62. After depletion of plastic-adherent and Fc+ cells by panning on dishes coated with normal serum, residual BM cells were cultured in gelatin coated flasks using murine rGM-CSF supplemented medium. Prior to analysis, non-adherent cells were re-depleted of contaminating Fc+ cells. Propagation of DC was monitored by double staining for FACS analysis (major histocompatibility complex (MHC) class II+ OX-62+, OX-19-). Functional assay, morphological analysis and evaluation of homing patterns of cultured cells revealed typical DC characteristics. MHC class II and OX-62 antigen expression increased with time in culture and correlated with allostimulatory ability. DC yield increased until day 7, when 3.3 × 106 DC were obtained from an initial 3 × 108 unfractionated BM cells. Significant numbers of DC can be generated from rat BM using these simple methods. This should permit analysis and manipulation of rat DC functions in vivo and in vitro. © 1995.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Chen-Woan, M
Delaney, CP
Fournier, V
Wakizaka, Y
Murase, N
Fung, J
Starzl, TEtes11@pitt.eduTES11
Demetris, AJ
Centers: Other Centers, Institutes, Offices, or Units > Thomas E. Starzl Transplantation Institute
Date: 27 January 1995
Date Type: Publication
Journal or Publication Title: Journal of Immunological Methods
Volume: 178
Number: 2
Page Range: 157 - 171
DOI or Unique Handle: 10.1016/0022-1759(94)00253-s
Institution: University of Pittsburgh
Refereed: Yes
ISSN: 0022-1759
Other ID: uls-drl:31735062126911, Starzl CV No. 1781
Date Deposited: 08 Apr 2010 17:30
Last Modified: 13 Jun 2021 01:55
URI: http://d-scholarship.pitt.edu/id/eprint/5167

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