A new cell line, designated as ZJBIQ-Chsu-I, was initiated from the fat body of larval Chilo suppressalis (Walker) (Lepidoptera: Pyralididae) in TNM-FH insect medium containing 15% fetal bovine serum. The polygonal cells (65.6%) were predominant among various cell types, and the diameter range was from 12.63 to 22.50 μm. The cell line showed a typical lepidopteran chromosome pattern ranging from 108 to 136 chromosomes in the majority of the cells. The population doubling time (PDT) of the cell line at the 15th passage was 62 h. This cell line was found to be susceptible to Spodoptera exigua nuclear polyhedrosis virus (SeNPV). By the DNA amplification fingerprinting polymerase chain reaction (DAF-PCR) technique, it was confirmed that cell line ZJBIQ-Chsu-I really originated from C. suppressalis.