The invention discloses a three-dimensional porous
chitosan / gelatine
microsphere, a preparation method thereof and an application thereof in liver
cell culture. The preparation method thereof comprises the following steps of: rapidly freezing mixture into spheres by dropping
liquid nitrogen by virtue of a high-pressure pulse microcapsule moulding instrument after
chitosan and gelatine B are mixed, carrying out secondary freeze-
drying after microspheres subjected to primary freeze-
drying are crosslinked and cured with saturated tripolyphosphate-85%
ethanol solution, fully hydrating the microspheres obtained by the secondary freeze-
drying, adding cross linking agent
carbodiimide / N-hydroxy succinyl, modifying the microspheres with gelatine A, removing unreacted
carbodiimide / N-hydroxy succinyl and gelatine A by washing after water bath and reaction in dark place are carried out, and carrying out
tertiary freeze-drying, thus the three-dimensional porous
chitosan / gelatine microspheres with the
diameter of 300-800Mum and the surface aperture of 50-200mu m are obtained, and the surface of each
microsphere is connected with the aperture of the interior of the
microsphere. The three-dimensional porous chitosan / gelatine microspheres obtained by the invention can be used for
in vitro culture of high-density and high-activity liver cells.