Transplantation of inbred adipose‐derived stromal cells in rats with plasma gel containing fragmin/protamine microparticles and FGF‐2

Abstract

Fragmin/protamine microparticles (F/P MPs) have been used as a cell carrier for adipose-derived stromal cells (IR-ASCs) in inbred male Fisher 344 rats, and for preservation and controlled-release of fibroblast growth factor (FGF)-2 and various cytokines in inbred rat plasma (IRP)-DMEM (Dulbecco's modified Eagle's medium) gel. In this study, we investigated the capability of an IRP-DMEM gel containing F/P MPs and/or FGF-2, as a three-dimensional (3D)-culture, to expand IR-ASCs. We found that IR-ASCs grow faster under 3D-culture conditions in low IRP (3%)-DMEM gel containing F/P MPs and FGF-2 without any animal serum than those under 2D-culture in low inbred rat serum (3%)-DMEM with F/P MPs and FGF-2. About 0.3 mL of IR-ASCs (about 4,000,000 cells mL⁻¹) grown in IRP (6%)-DMEM gel containing F/P MPs and FGF-2 disappeared 8 days after subcutaneous injection in rats, suggesting that they are rapidly biodegradable. The number of large (diameter ≥200 μm or containing ≥100 erythrocytes), medium (diameter = 20-200 μm or containing 10-100 erythrocytes) and small (diameter ≤20 μm or containing 1-10 erythrocytes) capillaries after injection with IR-ASCs in an IRP-DMEM gel containing both F/P MPs and FGF-2, as well as the thickness of tissue granulation per microphotograph at the injected site, was significantly higher than those after injection with IR-ASCs in an IRP-DMEM gel containing either FGF-2 or F/P MPs. Thus, IRP-DMEM gel containing F/P MPs and FGF-2 are useful and safe IR-ASC carriers that facilitate cell proliferation, vascularization, and tissue granulation locally at injection sites.

Effective Wound Healing in Streptozotocin-Induced Diabetic Rats by Adipose-Derived Stromal Cell Transplantation in Plasma-Gel Containing Fragmin/Protamine Microparticles

“…On the other hand, a 3-dimensional culture system for IR-ASCs in vitro has been reported in low IRP (3%)-DMEM gel containing F/P MPs, allowing the complete elimination of animal serum such as fetal bovine serum. 28 Plasma is a natural source of GFs, with PDGFs, TGFs, FGFs, and VEGFs stored in the > granules of platelets. Platelets activated by factors such as thrombin or calcium release these GFs from > granules.…”

“…27 We previously prepared injectable gels including IR-ASCs derived from inbred male Fisher 344 rats using low inbred rat plasma (IRP) (3%)-DMEM gel containing F/P MPs, and the gels were directly injected into normal rats for cell transplantation. 28 Researchers have used streptozotocin (STZ) to create experimental diabetes because it is a simple, inexpensive, and readily available method. 29 The mechanism responsible for impaired wound healing in STZ-induced diabetic rats (STZ-rats) is not completely understood.…”

Injectable Biomaterials in Plastic and Reconstructive Surgery: A Review of the Current Status

“…Sumi et al [117] loaded ADSCs into plasma gel with fragmin/protamine microparticles (F/P MPs) and FGF-2 in inbred male Fisher rats. The results indicated that the ADSCs grew faster under 3D-culture conditions in low plasma gel with F/P MPs and FGF-2 in vitro.…”

“…Black, dark gray, light gray, and white bars show gel containing both F/P MPs and FGF-2, F/P MPs, FGF-2, and none, respectively. Reproduced with permission[117] of John Wiley and Sons, Inc.…”

Biomedical Application of Low Molecular Weight Heparin/Protamine Nano/Micro Particles as Cell- and Growth Factor-Carriers and Coating Matrix

“…Various biomaterials have been used as cell carriers during cell implantations [ 57 , 58 ]. Those materials should fulfill several preconditions such as biocompatibility, biodegradability, low cytotoxicity and high affinity for the biological surface [ 57 , 58 ]. The biomaterial tested in this study, inbred rat plasma (IRP)-DMEM gel with LMWH/P N/MPs, can carry many IR-ADSCs and also act as a cell carrier in which the cells can grow.…”