Immobilization, stabilization, and activation of human stem cell factor (SCF) on fragmin/protamine microparticle (F/P MP)-coated plates.
Fragmin (low-molecular-weight heparin)/protamine microparticles (F/P MPs) immobilize to culture plates, thereby retaining the binding of heparin-binding cytokines such as human stem cell factor (SCF). The purpose of this study was to evaluate the ability of F/P MP-coating to immobilize, stabilize, and enhance SCF-activity. Cell assays showed that SCF and preimmobilized SCF on F/P MP-coated plates significantly stimulated the proliferation of human erythroleukemia cell line TF-1 in a concentration-dependent manner. Heparin and fragmin enhanced SCF-induced proliferation of chlorate-treated TF-1 cells, in which the biosynthesis of endogenous sulfated polysaccharides was blocked, on noncoated plates at a range of concentrations (2-8 microg/mL). However, heparin and fragmin had no effect on SCF-induced proliferation of chlorate-treated TF-1 cells on F/P MP-coated plates. The interaction of SCF with fragmin and F/P MPs prolonged the half-life of SCF bioactivity, and immobilized and protected SCF from inactivation, such as from heat and proteolysis. These results suggest that F/P MP-coated plates protect SCF and enhance its activity, and F/P MP-coating provides an excellent biomaterial to immobilize and retain heparin-binding cytokines, including SCF, in bioactive form for optimal expansion of hematopoietic cells.
Kishimoto
Satoko
S
Division of Biomedical Engineering, Research Institute, National Defense Medical College, Tokorozawa, Saitama, Japan.
Oonuma
Fumie
F
Nakamura
Shingo
S
Hattori
Hidemi
H
Nakamura
Shin-ichiro
S
Mori
Yasutaka
Y
Tanaka
Yoshihiro
Y
Harada
Yasuji
Y
Tagawa
Masahiro
M
Ishihara
Masayuki
M
D002455:Cell Division; D045744:Cell Line, Tumor; D017985:Dalteparin / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D011479:Protamines / Q000737:chemistry*; D019089:Stem Cell Factor / Q000502:physiology*
D006025:Glycosaminoglycans; D011479:Protamines; D019089:Stem Cell Factor; D017985:Dalteparin