Abstract: Three-dimensional cell culture is widely used in various fields of biomedical research. The research of three-dimensional culture scaffold materials is an important aspect. Nanocellulose with a length of 600-800 nm and a diameter of about 50-150 nm was prepared from bacterial cellulose by acidic hydrolyzing. The nanobacterial cellulose was combined with sodium alginate and Matrigel to prepare a three-dimensional culture scaffold to test its effect on MDCK cells′ three-dimensional growth. It was found that MDCK cells grew poorly in the gel scaffold only containing sodium alginate. A scaffold combining nanobacterial cellulose and sodium alginate could increase the growth of MDCK cells and form more cell clusters. Fluorescent staining nucleus and cell membrane of the cell cluster formed in the composite scaffold showed the MDCK cell mass exhibited a cystic structure unique to three-dimensional growth. This indicated that the nanobacterial cellulose and sodium alginate complex scaffold could be used for three-dimensional culture of MDCK cells. Adding 0.5% Matrigel to the complex significantly promoted the three-dimensional growth of MDCK cells. This method of adding a small amount of Matrigel to the complex to supplement the extracellular matrix necessary for three-dimensional growth of cells had certain prospects in application development. The research would provide a preliminary basis for exploring nanobacterial cellulose as a scaffold material for three-dimensional cell culture.

Key words: three-dimensional culture, scaffold material, nanobacterial cellulose, sodium alginate