关键词: MRC-5细胞, 工作细胞库, 传代稳定性, 狂犬病疫苗

Abstract:

To establish a working cell bank of medical research council 5 （MRC-5） cells and conduct relevant research on the application of this cell bank in the production of inactivated rabies vaccine， the MRC-5 working cell bank was established in strict accordance with the Pharmacopoeia of the People's Republic of China， and all mandatory tests were performed following the pharmacopoeial specifications. Meanwhile， the growth curve， passage stability， and rabies virus susceptibility of MRC-5 cells were systematically investigated. The cells from the established MRC-5 working cell bank exhibited a typical long spindle shape with clear edges and distinct outlines， and the cell viability reached 99.03%. The results of sterility test， mycoplasma test， and exogenous factor test all conformed to the pharmacopoeial requirements. The growth curve of MRC-5 cells presented a standard sigmoidal （S） shape， with cells entering the logarithmic growth phase in 2~3 days and a cell doubling time of 21 hours. Moreover， the cell morphology remained normal even after continuous subculture to the 66th passage. After inoculation of the PM strain of rabies virus into MRC-5 cells of 36， 38， 40 and 42 passages， the mean virus titers were determined as 6.57±0.21， 6.40±0.26， 6.23±0.21 and 6.40±0.17 lgCCID??·mL^-1， with the coefficient of variation （CV） ranging from 2.70% to 4.13%. In conclusion， the established MRC-5 working cell bank meets the requirements specified in the Pharmacopoeia of the People's Republic of China. It shows excellent passage stability and high susceptibility to rabies virus， indicating that it is suitable for the industrial production of inactivated rabies vaccine.

Key words: MRC-5 cells, working cell bank, passage stability, rabies virus vaccine