Screening and Application ofHEp-2 Cell Culture Medium without Ingredients of Animal Origin

Indirect immunofluorescence is a commonly used method for preliminary diagnosis of autoimmune diseases, which has the characteristics of simplicity and high specificity. Human laryngeal cancer cells (HEp-2 cells) are the main raw material for preparing indirect immunofluorescence detection kits. Driven by the international trend of human serum free, countries such as the United States and European Union has stopped using serum in biopharmaceuticals in 2010. Henceforth, the development of culture media serum-free, and even animal free, for HEp-2 cells is an urgent task. In the study, 4 different serum free culture media were used for adaptive culture and domestication of Hep-2 cells, pancreatin of animal original was replaced by recombinant trypsin during cell isolation, and the differences between 3 serum -free/non animal original culture media and normal serum-containing media were compared through cell counting and cell morphology. Besides, differently cultured cell sheets were used during indirect immunofluorescence test, results showed that cell doubling duration in serum 1640 culture media was 25 hours and cell doubling duration in CD-100 and OPM-AM146 culture media was 28 hours, indicating no obvious difference compared with control group cultured in serum media concerning doubling duration and proliferation curve; cell doubling duration in ToCell® Vero CD culture media was between 32-37 hours, representing a significant difference in doubling time. And there is no difference in the detection limit for known positive samples during the indirect immunofluorescence assay performed using cells cultured in the screened culture media.