Induction of the murine "W phenotype" in long-term cultures of human cord blood cells by c-kit antisense oligomers.
The murine white (W) spotting locus is the site of the c-kit gene and encodes a tyrosine kinase receptor while the complementary Steel (Sl) locus encodes its ligand. Mutations at either locus have profound effects on hematopoiesis, particularly erythroid and mast cell proliferation. We added c-kit antisense oligonucleotides to long-term suspension cultures of enriched human umbilical cord progenitor cells. This resulted in the suppression of c-kit gene expression and the preferential suppression of the generation of erythroid burst-forming cells (BFU-E) which extended over the life of the culture (3 weeks). The results provide an in vitro model of the "W phenotype" in human hematopoiesis and confirm the importance of c-kit gene function in early erythropoiesis. Because the generation of BFU-E was suppressed even after c-kit gene expression had recovered, this gene product may be critical to the erythroid commitment process.
Migliaccio
A R
AR
Lindsley F. Kimball Research Institute, New York Blood Center, New York 10021.
Migliaccio
G
G
Mancini
G
G
Ratajczak
M
M
Gewirtz
A M
AM
Adamson
J W
JW
D000818:Animals; D016375:Antisense Elements (Genetics) / Q000235:genetics; D001483:Base Sequence; D001773:Blood Cells / Q000502:physiology*; D002452:Cell Count / Q000187:drug effects; D002478:Cells, Cultured; D005312:Fetal Blood; D005786:Gene Expression Regulation / Q000187:drug effects*; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000187:drug effects; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D051379:Mice / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010641:Phenotype; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D019009:Proto-Oncogene Proteins c-kit; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D016184:Receptors, Colony-Stimulating Factor / Q000235:genetics*; D019089:Stem Cell Factor; D013997:Time Factors
D016375:Antisense Elements (Genetics); D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3; D015345:Oligonucleotide Probes; D016376:Oligonucleotides, Antisense; D011518:Proto-Oncogene Proteins; D016184:Receptors, Colony-Stimulating Factor; D019089:Stem Cell Factor; D019009:Proto-Oncogene Proteins c-kit; D020794:Receptor Protein-Tyrosine Kinases