Research For The Technology Of Setting Up Insect Cells Bank And The Primary Cell Culture For Insect Cells

According to the situation that there are only more than twenty insect ceils linesat present in our country, we set up the first professional insect cells bank byourselves as the same time collecting the insect cells lines in and abroad, which hasprovided the necessary technology and the material store for the biotechnology,molecular biology, biochemistry and genetics in our country. Different tissues ofmany sorts of insects were tried to be cultured in vitro in this research, whichprovided the necessary basic theory and useful reference for setting up the newinsect cells lines. We also studied the technology of building the insect cells banksystematically in the experiment. It provided important academic theory for theresearch of insect cells. 1. The primary research for the technology of building insect cells bank We statistically observed the seven insect cell lines in cell morphology, whichwere fetched from Japan. After we drew the growth curve of the seven insect cellslines, counted the Population Doubling Time (PDT), and compared the results ofthe cryopreservation and thaw of the insect cells, we drew the conclusions as follows:? The cell lines FRI-SPIM-1229 came from five instar larva's fat bodies of lipocyte ofSpilosoma imparilis Butler. The Population Doubling Time (PDT) was 87.8h;? The cell lines Grace's A.e cell came from Aedes albopictus Skuse whose Population Doubling Time (PDT) was 62.6h;? The cell lines NIAS-MaBr-85 came from five instar larva's fat bodies of lipocyte of Mamestra brassicae Linnaeus whose Population Doubling Time (PDT) was 50.23h;? The cell lines NIAS-Px-58 came from ovary of the pupae of Papilio xuthus Linnaeus whose Population Doubling Time (PDT) was 63.5h;? The cell lines NIH-SaPe-4 came from embryos of imago ovary of Sarcophaga peregrina Sopoli whose Population Doubling Tune (PDT) was 92.9h;? The cell lines SES-MaBr-2 came from larva's fat bodies of Mamestra brassicae Linnaeus whose Population Doubling Time (PDT) was 135h;? The cell lines NLAS-MaBr-92 came from five instar larva's blood of Mamestra brassicae Linnaeus whose Population Doubling Time (PDT) was 45.7h.Besides mensurating the growing and morphologic characteristic of the seveninsect cell lines, we tried to compare. That is to say that we chose the cryogenicrefrigerator and the liquid nitrogen vat, took the dimethylsulfoxide and glycerin astheprotectant, and cryopreserved the seven insect cell lines. In the experiment wecompared cryopreservation and anabiosis of the seven insect cell lines, and discussedand summed up some questions that needed to be paid attention to in preserving thecells in building the insect cells bank. And then we got the necessary reference datafor setting up the insect cells bank. Also we made it become the basic theory of thebuilding technology of the insect cells bank. 2. The primary cell culture for insect cells(1) The primary cell culture for egg aud larva tissues of Dendrolimnskikuchii Matsumura This was the first time that people cultured the egg and larva tissues ofDendrolimns kikuchii Matsumura of Yunnan in vitro in and out of China. After thecell was moved into the culture flask, on the 3rd day, we could observe that the cellsmoved out of the tissues. The cells were spindle-shaped and adhered to the wall ofthe culture flask. But the cells stopped growing on the 10th day and died on the 55thday. (2) The primary cell culture for the blood-lymph of Blaps japanesis yunnanensis Mars This was the first time to culture Coleoptera cells in vitro in China. In this research we cultured in vitro the blood-lymph of the imago Blaps japanesis yunnanensis Mars which was the pharmaceutic insect in Yunnan. There were cells adhering to the wall of culture flask after the cells were moved into the culture flask for 3 days. On the 4th day, there were new cells moved out of the tissues, and most of the ce...