蓖麻蚕蛹卵巢细胞株的建立及其对核型多角体病毒敏感性的研究

ESTABLISHMENT OF A CELL LINE FROM PUPAL OVARY OF PHILOSAMIA CYNTHIA RICINI AND ITS SUSCEPTIBILITY TO NUCLEAR POLYHEDROSIS VIRUS

用蓖麻蚕（Ｐｈｉｌｏｓａｍｉａｃｙｎｔｈｉａｒｉｃｉｎｉ）雌蛹卵巢组织进行离体细胞培养，建立了细胞株，命名为ＷＶＢ－Ｐｃｒ－０２．在建株过程中，原代培养一个月左右，就开始传代，细胞生长稳定，分裂比值为１∶２，每４～５ｄ传代一次．细胞主要为贴壁的圆形细胞，其次是梭形细胞．细胞群体倍增时间为６０～６４ｈ．观察细胞染色体，其二倍体蚕的染色体为５６条，建株后染色体数为１００～１１０条，与其他蚕细胞类似．用同源蓖麻蚕核形多角体病毒感染后，观察了细胞病理变化及病毒在蓖麻蚕细胞中的复制．细胞感染率可达９０％以上，多角体产量为２．４×１０７ＰＩＢ·Ｌ－１．

A cell line was established from pupal ovary of Philosamia cynthia ricini (Pcr) and was designatet as WVB Pcr 02. The cell line WVB Pcr 02 was established from pupal ovarian tissues of Philosamia Cynthia ricini . The cell line has been transfered for 102 passages in 36 months. The medium BML TC100 supplemented with 20% Fetal calf serum was suitable for culturing pupal ovaries of Philosamia Cynthia ricini and Tc Tryptose broth has the ability to stimulate the growth of the silkworm cells.It showed the same character as other cell of silkworm in the process of growth. suspension containing cell masses and single cells was subcultured with the split ratio of 1∶2.The cells population doubling time was 60～64 h and the maximum cell population per milliliter was 4.3 times as the initiated cell density. The homologous PcrNpv was used to infect cell line. Polyhedral inclusion bodies formed in the nuclei of the WVB Pcr 02 cell line, which was susceptible to the virus. Ovary 90% of cells were infected. Product dosages of 2.4×10 7 PIB/mL. When viewed with an electron microscope, the infected cells at five days post inoculation showed viral matrix in their nuclei. Nucleocapsids were formed in the matrix and were enveloped multiply as bundles of virus rods. This cell line was found to susceptible Pcr NPV. [WT5HZ〗