Fibrillar collagen assembled in the presence of glycosaminoglycans to constitute bioartificial stem cell niches in vitro

Abstract

Fibrillar collagen was reconstituted from mixtures of monomeric tropocollagen and heparin or hyaluronic acid, respectively. Turbidity measurements were utilized to follow the fibrillar assembly and demonstrated the influence of the concentration of the glycosaminoglycan on the maximum optical densities. Thin film coatings of maleic anhydride copolymers were utilized for the covalent immobilization of the fibrillar assemblies to solid supports. Quantification of surface-bound collagen was accomplished by ellipsometry and HPLC-based amino acid analysis indicating that less collagen was immobilized in the presence of the glycosaminoglycans. SEM and AFM revealed various sizes and shapes of the immobilized fibrillar assemblies if collagen fibrils were prepared in the presence of heparin or hyaluronic acid. Human hematopoietic stem cells (HSCs) were cultivated on the surface-bound collagen fibrils and the migration of adherent cells was studied by time-lapse microscopy. Migration rates on fibrillar structures were significantly lower then on tropocollagen indicating a more intimate contact of HSCs to the fibrillar substrates.