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Involvement of CSF‐1 in generating a stroma‐independent hematopoietic stem cell line
Author(s) -
Heberlein Christoph,
Friel Jutta,
Itoh Katsuhiko,
Medlock Eugene,
Li Luke,
Nakayama Naoki,
Stocking Carol,
Geldmacher Maren,
Ostertag Wolfram
Publication year - 2006
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.20505
Subject(s) - stroma , haematopoiesis , stem cell , line (geometry) , hematopoietic stem cell , microbiology and biotechnology , biology , cell culture , cancer research , immunology , genetics , mathematics , immunohistochemistry , geometry
Abstract The hematopoietic stem cell line, Myl‐D7, is maintained by a self‐renewing stem cell population that spontaneously generates myeloid, lymphoid, and erythroid progeny. MS‐5 stromal cells are necessary for the growth of Myl‐D7 cells. One component of the Myl‐D7 cells proliferation activity released by MS‐5 stromal cells was enriched by Q sepharose fractionation and shown to be colony stimulating factor‐1 (CSF‐1) by Western blotting, BAC1.2F5 cell bioassay and inhibition of Myl‐D7 proliferation by CSF‐1 antibody. The requirement of Myl‐D7 cells for CSF‐1 was also demonstrated independently by selecting for rare, stroma‐independent Myl‐D7 mutant clones able to grow without stroma and additional factors. Eighty‐nine stroma‐independent mutant clones were obtained and belonged to two classes. The majority of mutants did not secrete any growth promoting activity. The second, rarer class of mutants releases a factor that stimulates proliferation/survival for up to several months and approximately half of the secretors express high levels of CSF‐1 mRNA. Wild type Myl‐D7 grown with supernatants from the secretor cells retained the stem cell phenotype. These data suggest that CSF‐1 may act as a key factor in stroma‐regulated hematopoiesis and cell–cell interaction. J. Cell. Physiol. 206: 556–562, 2006. © 2005 Wiley‐Liss, Inc.