The Behavior of Embryonic Neural Cells within the 3D Micro-structured Collagen-Based Scaffolds

Abstract

Synthesis of scaffolds providing mechanical support for the growing cells is important in reconstruction of the tissue. Addition of chondroitin sulfate (CS) to collagen scaffolds was proved to regulate the neural cells growth and differentiation. The aim of the study is to test whether collagen-CS cross-linked scaffolds could be used for embryonic neuronal cell culture. Embryonic neuronal cells were isolated from rat foetus brains and their differentiation into neurons or astrocytes was characterized by flow cytometry. Then, the entrapment and distribution of cells within collagen alone or composite collagen with CS scaffolds was studied. The cells were applied to the scaffold and stained with bisbenzimide and then counted. Finally the MTT test was performed. The results suggest that embryonic neuronal cells were differentiated into neurons (MAP2 positive cells) or astrocytes (GFAP positive cells). The cells entered into the two tested 3D scaffolds; however their distribution in the tested scaffolds was not homogenous. The MTT test showed more intensive metabolism of the cells in collagen with CS scaffolds comparing with controls (cells seeded on laminin) or cells growing in scaffolds composed only from cross-linked collagen. Conclusion: the both collagen and collagen with CS scaffolds are the good carriers for the embryonic neuronal cells. However, the sample composed of collagen with CS constitutes better conditions for embryonic neuronal cells culture.