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Differentiation of Human Hematopoietic Cells Increases Expression of a Gene Transferred by a Retroviral Vector
Author(s) -
Holland Christie A.,
Chada Sunil,
Wright Jocyndra,
Whitney Constance,
Harigaya Kenichi,
Greenberger Joel S.,
Newburger Peter E.
Publication year - 1989
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.46.3.221
Subject(s) - biology , haematopoiesis , microbiology and biotechnology , cell culture , stromal cell , viral vector , hl60 , gene expression , k562 cells , northern blot , neomycin , vector (molecular biology) , gene , stem cell , cancer research , genetics , recombinant dna , antibiotics
To study expression of a retroviral vector in human hematopoietic lineages, two established human hematopoietic cell lines (HL60 and K562) and a human adherent stromal cell line (KM101) were infected with the vector pZIP‐SV(X). Expression of the transferred neomycin resistance gene (neo) of pZlP‐SV(X) was evaluated as the ability of the cells to form colonies (>50 cells) in an agar assay in the presence of the neomycin analogue, G418. After infection, all three cell lines produced colonies resistant to G418. The level of neo mRNA in separate colonies was analyzed by Northern blot analysis. The neo gene transferred by the vector pZIP‐SV(X) was expressed in both human hematopoietic and stromal cell lines. In addition, primary adherent human stromal cells infected with pZIP‐SV(X) grew in the presence of G418. To determine if differentiation of hematopoietic cells affects expression of the retroviral vector, HL60 cells infected with pZIP‐SV(X) were induced to differentiate, and the level of neo mRNA measured. The amount of neo mRNA increased when HL60 cells were induced to differentiate along the granulocytic pathway. Conversely, when HL60 cells were induced toward monocytoid differentiation (TPA), the level of neo mRNA did not significantly increase. We conclude that the neo gene transferred by a retroviral vector, pZIP‐SV(X), is functionally expressed. In addition, expression of the transferred neo gene is regulated during myeloid differentiation of HL80 cells.