CX-2 Microcarrier Preparation And The Characteristics In Cell Culture

The cultivation on microcarriers (MCs) is a perfect method on the large-scale animal cell culture, which is the complete combination of adhesive culture and suspending culture. In this system the properties of MCs are critical effect to cell culture, in fact, there is still no one perfect MC in all-present commercial products. Therefore, it is necessary to develop superior quality MCs. In this research, a new MC, CX-2 has been developed using Carboxymethyl-chitosan (CM-CH). In this paper, the relationship on inherent viscosity, degree of deacetylation and degree of carboxymethyl has been studied; the preparation process of CX-2, the histocompatibility and the animal cell culture have been described. The results provide foundational theories for preparation of the MCs and application in large-scale animal cell culture.CM-CH is a water-soluble derivative of chitosan. The results manifest that it is advisable to prepare certain molecular weight CM-CH from corresponding MW CH; Deacetylation degree and carboxymethyl degree of CH are unchanged after a series of reactions. The optimal raw material of CM-CH to prepare CX-2 is: viscosity 260ml/g, deacetylation degree more than 90 per, andcarboxymethyl degree more than 85 per. The physicochemical properties of CX-2 correspond to Cytodex-3 MC produced by Pharmacia, including of color, loss of drying, area, density, size, soakage, dilation, stability, size distribution, mechanical strength, surface structure, etc., but its cost of manufacture is lower than Cytodex-3 's.The results suggest that CX-2 can remarkably adsorb serum proteins on comprehensive pH conditions and has good biocompatibility. Some animal cells such as Vero, BHK and BB, could adhere better to CX-2 MC. The cells proliferate quickly on CX-2 MC from 24 to 72 hours in cell culture, after cell culture 120 hours, there are multilayer cells growing on the CX-2 MC surfaces, and some cells can transfer from the surfaces of the cell-covered CX-2 MCs to new- added ones. All studies indicate that CX-2 is available in animal cell culture. It is viable that CX-2 MC can be used more times in cell culture.