Proliferation of 3T3 fibroblasts in a spinner type reactor on microcarriers of fibrin-alginate

Abstract

The microcarriers have become an alternative of cell culture in order to increase the contact area per unit volume of culture medium. The aim of this work is the fabrication of microcarriers from a mixture of sodium alginate and human blood plasma, to be used as a fibroblast support during proliferation in bioreactors. Microcarriers were obtained by spraying through a coaxial nozzle over calcium chloride. Poly-L-Lisine (PLL) and fibrinogen were adsorbed on the surface of microcarriers. The microcarriers were observed by atomic force microscopy (ATM) and scanning electronic microscopy (SEM). Adhesion of 3T3 fibroblasts was evaluated over the microcarriers surface. Microspheres between 150 μm and 500μm of diameter were obtained, it was evaluated the cellular adhesion and a first-order kinetics was adjusted with a constant k = 0.007 min-1. Celullar adhesion was observed on the microcarriers using confocal microscopy and fluorescence microscopy. Fibroblasts grown in the reactor showed that it is possible to reduce the doubling time from 23.4 until 16.3 h.