Genetic approach and phenotype-based complementation screening for identification of stroma cell-derived proteins involved in cell proliferation.
The functional capacities of stromal cell lines to support stem cell activity are heterogeneous and the mechanism of how they support bone marrow cultures remains unclear. Recently, we reported a strategy of functional analysis in which a genetic approach is combined with phenotype-based complementation screening to search for a novel secreted growth factor from mouse bone marrow stroma called ShIF that supported proliferation of bone marrow cells. To investigate the role of stromal cells in hemopoiesis, we extended this strategy to search for stroma-derived proteins that induce cell proliferation by establishing stroma-dependent Ba/F3 mutants of three stroma cell lines from two mouse tissues. Seven stroma-dependent Ba/F3 mutants were used as responder cells to identify cDNAs from stroma cell lines whose products supported proliferation not only to the mutant cells but also to hemopoietic progenitor cells in vitro.
Tulin
Edgardo E
EE
Chugai Research Institute for Molecular Medicine, Inc., 153-2 Nagai, Niihari, Ibaraki, 300-4101, Japan. tulin@cimmed.com
Onoda
Nobuhisa
N
Hasegawa
Masakazu
M
Nosaka
Tetsuya
T
Nomura
Hitoshi
H
Kitamura
Toshio
T
D000818:Animals; D002455:Cell Division / Q000235:genetics; D002460:Cell Line; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology; D015723:Gene Library; D006133:Growth Substances / Q000235:genetics; D007378:Interleukins; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D011506:Proteins / Q000235:genetics*; D017494:Proton Pumps; D006180:Proton-Translocating ATPases; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*; D014162:Transfection
C432920:Atp6v0a2 protein, mouse; D018076:DNA, Complementary; D006133:Growth Substances; D007378:Interleukins; C442281:Mydgf protein, mouse; D011506:Proteins; D017494:Proton Pumps; D006180:Proton-Translocating ATPases