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Isolation of rat bone marrow mast lineage cells using Thy 1.1 and rat stem cell factor
Author(s) -
Medlock Eugene S.,
Mineo Chrysa,
Housman Jerry M.,
Elliott Gary S.,
Trebasky Lisa D.,
Langley Keith E.,
Zsebo Krisztina M.
Publication year - 1992
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041530309
Subject(s) - bone marrow , stem cell factor , stem cell , isolation (microbiology) , biology , lineage (genetic) , mast cell , mast (botany) , microbiology and biotechnology , haematopoiesis , immunology , genetics , gene , bioinformatics
Recent reports have shown that various marrow‐derived cell populations respond vigorously to recombinant rat stem cell factor (rrSCF 164 ), one form of the kitligand. In the present study, we isolated cell populations from rat bone marrow using the Thy 1.1 antigen (an antigen that in the rat is differentially expressed on primitive hemopoietic progenitor cells) and fluorescently conjugated rrSCF 164 (rrSCF 164 ‐PE). We show that rrSCF 164 only stimulates cells that are enriched in the brightest Thy 1.1 populations (Thy 1.1 bright ). Numerous cell lines were generated by serial passage in rrSCF 164 containing medium, and the prototypic cell lines have been designated SRT002 and SRT003. Each cell line retains the Thy 1.1 bright phenotype and does not respond to interleukins (IL) 1–8, IL‐10, granulocyte (G) colony‐stimulating factor (CSF), granulocyte macrophage (GM) CSF, M‐CSF, or crude preparations of mitogen‐stimulated T‐cell supernatants. The Thy 1.1 bright population of rat marrow was subdivided into a subset that binds rrSCF 164 ‐PE (Thy 1.1 bright , rrSCF 164+ ). The majority of these cells possess certain characteristics in common with marrow‐derived mast cells and the Thy 1.1 bright , rrSCF 164 responsive cell lines, having similar granule morphology, being metachromatic, and reacting positively with alcian blue. Moreover, rats treated with rrSCF 164 displayed significant increases in Thy 1.1 bright , rrSCF 164+ cells in the bone marrow. These studies show that the combination of Thy 1.1 and rrSCF 164 makes possible the isolation of a unique subset of rat bone marrow cells that differentially express the Thy 1.1 antigen and the cell surface receptor c‐ kit , the majority of which are morphologically similar to marrow‐derived mast cells. © 1992 Wiley‐Liss, Inc.