Cytokine-dependent proliferation of human CD34+ progenitor cells in the absence of serum is suppressed by their progeny's production of serine proteinases.
In this study, we demonstrate that the synthesis and release of serine proteinases by hematopoietic cells affects the in vitro proliferation of hematopoietic progenitor cells (HPCs) in response to proteins, including hematopoietic growth factors (HGFs), transferrin, insulin, and albumin in serum-free cultures. In serum-free cultures, bone marrow mononuclear cells or the CD34- progeny of the CD34+ cells were shown to release the serine proteinases human neutrophil elastase (HNE), cathepsin G (Cath G), and proteinase 3 (Pr3). In the absence of serum, we showed that HNE, Cath G, and Pr3 rapidly and dose-dependently degraded HGF and other proteins present in the medium, resulting in decreased proliferation of HPCs. Addition of the serine proteinase inhibitors alpha1-proteinase inhibitor (alpha1-PI) or the secretory leukocyte proteinase inhibitor (SLPI), but not leupeptin, aprotinin, or AEBSF (4-[2-aminoethyl]-benzenesulfonylfluoride hydrochloride), could completely prevent the degradation of proteins relevant to the growth of hematopoietic cells. Thus, the addition of serine proteinase inhibitors like alpha1-PI or SLPI may be critical for the expansion of CD34+ cells or gene transfer into CD34+ cells or other hematopoietic cells in vitro using serum-free media under good manufacturing practice conditions.
Goselink
Henriette M
HM
Laboratory of Experimental Hematology, Department of Hematology, C2-R, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands. H.M.Goselink@lumc.nl
Hiemstra
Pieter S
PS
van Noort
Peter
P
Barge
Renée M Y
RM
Willemze
Roel
R
Falkenburg
J H Frederik
JH
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000201:enzymology / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects*; D016207:Cytokines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006412:Hematopoietic Stem Cells / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006801:Humans; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013997:Time Factors
D018952:Antigens, CD34; D016207:Cytokines; C066855:HGF protein, human; D015842:Serine Proteinase Inhibitors; D017228:Hepatocyte Growth Factor; D012697:Serine Endopeptidases