Fragmin/Protamine Microparticle‐Coated Matrix Immobilized Cytokines to Stimulate Various Cell Proliferations With Low Serum Media

Abstract

Fragmin/protamine microparticles (F/P MPs) have been shown to bind to culture plates, thereby retaining heparin-binding cytokines. Most protocols for in vitro cultures of human microvascular endothelial cells (hMVECs), human dermal fibroblast cells (hDFCs), and hematopoietic cell line (TF-1) include high fetal bovine serum (FBS) (10%) medium as a nutritional supplement. Growth rates of those cells on the F/P MP-coated plates were higher in low FBS (1%) medium containing fibroblast growth factor (FGF)-2 (for hMVECs and hDFCs) and interleukin (IL)-3/granulocyte-macrophage colony-stimulating factor (for TF-1 cells) than without coating. The cytokines in low FBS medium were shown to be immobilized on the F/P MP-coated plate and released into the culture medium with a half releasing time of 4-5 days. Furthermore, those cells grew well on each cytokine-preimmobilized F/P MP-coated plate in low FBS medium. Thus, the F/P MP-coated matrix with adequate heparin-binding cytokines may provide biomaterials for controlling cellular growth and differentiation.

PRP&F/P MPs Improved Survival of Dorsal Paired Pedicle Skin Flaps in Rats

“…F/P MPs were able to protect activity of GFs in PRP from inactivation by incubation at 37 C for more than 7 d [19]. A local injection of PRP&F/P MPs showed substantial induction of vascularization and fibrous tissue formations by absorbing, stabilizing, and gradually releasing heparin-binding GFs from PRP&F/P MPs [23,24]. Those results indicated that GFs in the activated PRP are released from a-granules and bound to and stabilized on F/P MPs, and that GFs incorporated onto F/P MPs are gradually diffused and released upon biodegradation of F/P MPs in vivo.…”

Culture on a Fragmin/protamine-Coated Plate Suppresses the Collagen Type IαI and TGF-β1 mRNA Expression of Rat Hepatic Stellate RI-T Cells

“…TGF-β1 and PDGF are absorbed onto the membrane surface of fibroblasts through the ECM, which inhibits binding to each receptor [15]. F/P-MPs can also absorb TGF-β1 and HGF [14]. The RI-T cells used in the present study express TGF-β1 mRNA.…”

“…The supernatant was removed, and the solution of F/P-MPs was filled to 1 ml with PBS. More than 7 mg of dry F/P-MPs was obtained from 1 ml of F/PMPs solution [14].…”

“…Kishimoto et al [14] reported that microparticles that are a mixture of Fragmin ® (dalteparin; a low-molecular-weight heparin with anti-coagulant properties), and protamine, an antagonist of heparin, have ECM-like functions in fibroblast culture systems. Fragmin is one of the heparan sulfate proteoglycans; it charges negatively with sulfate group [9] and interacts with a variety of functional proteins such as cytokines [11].…”

Fragmin/Protamine Microparticles (F/P MPs) as Cell Carriers Enhance the Formation and Growth of Tumors In Vivo

“…In addition, it has been demonstrated that various GFs (including FGFs, HGF, VEGFs, TGFs, and PDGFs) are effectively immobilized on the F/P MPs. 6,7 The present study evaluates F/P MPs as cell carriers that stimulate the growth of 3LL, B16, and Huh7 tumors in vivo. The interaction of these cells with F/P MPs induced the in vitro formation of tumor cell/F/P MP aggregates, and the cells remained viable in suspension.…”