Modulation of human cord blood progenitor cell growth by recombinant human interleukin 3 (IL-3), IL-6, granulocyte-macrophage colony stimulating factor (GM-CSF) and stem cell factor (SCF) in serum-supplemented and serum-free medium.
We evaluated the growth of cord blood myeloid progenitors or colony forming units granulocyte-macrophage (CFU-GM) and their response to various recombinant growth factors or colony stimulating factors (CSFs): interleukin 3 (IL-3), IL-6, granulocyte-macrophage CSF (GM-CSF) and stem cell factor (SCF). Using classical stimulant (human placenta conditioned medium or HPCM), we observed a significantly higher day-14/day-7 CFU-GM ratio in CB than in bone marrow (BM). The association of IL-3, IL-6, GM-CSF and SCF induced significantly more CB day-14 CFU-GM than HPCM. This effect is significantly greater in CB than in bone marrow. Since fetal calf serum (FCS) is known to contain inhibitors, we have compared the ability of CSFs to induce CFU-GM formation in FCS-supplemented and FCS-free culture. In CB, using HPCM, we obtained significantly more CFU-GM in FCS-free medium than in FCS-supplemented medium. This difference was corrected by the addition of anti-transforming growth factor-beta (TGF-beta) neutralizing antibody. However, with the association of the four CSFs, no significant difference between FCS and FCS-free culture was observed.


CONCLUSIONS
a) day-14/day-7 CFU-GM ratio was higher in CB than in BM indicating that CB CFU-GM are more primitive than BM CFU-GM; b) FCS can be successfully replaced by serum-free medium; c) FCS contains inhibitors of day-14 CFU-GM and among them TGF-beta; and d) the association IL-3, SCF, GM-CSF and IL-6 seems able to totally overcome the inhibitory effect of FCS.
De Bruyn
C
C
Service de Médecine Interne et Laboratoire d'Investigation Clinique Henri Tagnon, Institut J. Bordet, Brussels, Belgium.
Delforge
A
A
Bron
D
D
Ley
P
P
de Hemptinne
D
D
Stryckmans
P
P
D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D003114:Colony-Forming Units Assay; D003470:Culture Media; D005312:Fetal Blood / Q000166:cytology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology*; D006410:Hematopoiesis / Q000187:drug effects / Q000502:physiology; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D066298:In Vitro Techniques; D007231:Infant, Newborn; D007377:Interleukin-3 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D019089:Stem Cell Factor; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000502:physiology
D003470:Culture Media; D006133:Growth Substances; D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3; D015850:Interleukin-6; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D016212:Transforming Growth Factor beta; D016178:Granulocyte-Macrophage Colony-Stimulating Factor