Patent ID: 6194210
Filing Date: 2001-02-27
Classification: A61K,C12N,Y02A

Abstract:
A method for producing large quantities of hepatitis A virus (HAV) in an aggregated microcarrier-cell culture which comprises retaining an HAV-infected MRC-5 cell population in an aggregated and attached state, with no observable floating MRC-5 cells due to cell sloughing from the microcarriers, comprising:a) planting MRC-5 cells in a suitable culture medium at a planting concentration sufficient to achieve uniform attachment of the cells to glass coated microcarriers, such that essentially every microcarrier has at least one cell attached,b) growing the cells so attached to the microcarriers in a culturing vessel using a rate of agitation equal to the critical off bottom suspension rate,c) infecting the microcarrier aggregates in late log phase or early stationary phase with HAV, wherein said MRC-5 cells remain in an attached state, with no observable floating cells due to cell sloughing from the microcarriers,d) culturing the infected microcarrier-cell aggregates, ande) harvesting and recovering the HAV so produced.