1株悬浮培养的Vero细胞驯化及其生物学特性研究OACSTPCD

Objective To domesticate adherently cultured Vero cells into suspension-cultured cells and investigate their biological characteristics.Methods Cell culture was optimized using serum-free media and the"shaker adaptation method."Additionally,tumorigenicity,short tandem repeat(STR)profiles,morphological characters under transmission electron microscope(TEM),and influenza virus sensitivity were analyzed in these cells.Results Suspension cell line Vero-S was successfully established.The growth period of Vero-S cells ranged from 24 to 72 hours,with a plateau phase between 108 and 144 hours,and a decline phase after 144 hours.The population doubling time(PDT)was 36 hours,and cell density ranged from 14.01 to 14.88×108 cells/L.Average cell roundness ranged from 0.73 to 0.75,and cell diameter ranged from 12.12 to 14.44 µm.Vero-S cells formed tumors subcutaneously in nude mice,while Vero cells before suspension culture did not exhibit tumorigenicity.STR analysis indicated a primate origin of the cell line.Under electron microscopy,cell structure appeared intact.Vero-S cells showed insensitivity to influenza virus infection.Conclusion The established suspension cell line Vero-S exhibiting growth patterns similar to adherent cells,with uniform cell size and high-density growth supported by the established suspension culture medium.STR results demonstrated complete genetic stability of suspended Vero cells,with no apparent differences observed under transmission electron microscopy compared to adherent Vero cells.This study provides reference for suspension culture domestication techniques and can be applied in mechanistic studies of cell tumorigenicity.