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Insect cell culture technology is one of the main areas of cell engineering has developed rapidly in recent years, and are widely used in medicine, agriculture and biology. In this paper, the lepidopteran (Lepidoptera) dead leaves the Tortricidae (Lasiocampidae) dendrolimi are (Dendrolimus Germer) of Johnson pine caterpillars (Dendrolimus punctatus tehchangensis Tsai) test material, its blood lymphocytes, ovarian tissue, embryos, as well as the new hatch larval tissues cultured in vitro test. Meanwhile, the research and development of computer management system for insect cell library management, standardized and effective to achieve long-term insect cell library management laid the foundation. The main results are as follows: 1, the Johnson dendrolimi tissue cells cultured This study explored the the Johnson dendrolimi tissue in vitro culture conditions obtained a suitable culture methods for various organizations and cells in vitro proliferation and survival of a long time. (1) medium: By comparison, Grace, MGM-450, MM, TNM-FH insect medium applicable to the in Johnson dendrolimi blood lymphocytes, ovarian cells, embryonic cells, and the newly hatched larvae cell culture . (2) the Johnson dendrolimi larvae blood lymphocyte culture: standing method to collect blood lymphocytes, the initial density control to achieve better effect of culture between 0.27 × 106 ~ 0.53 × 106 cells / mL. Cultured cells by the circular suspended growth type, circular adherent growth type, fusiform adherent growth type, a polygonal adherent growth type cells and reticular cells, etc. of different types of cells. By changing fluid cultures, pine blood lymphocytes in vitro survival of more than 1 year. (3) the Johnson dendrolimi pupal ovarian cell culture: select the ovarioles end silk and reproductive area portion (about 5mm), trypsin digestion and mechanical shear method of combining discrete organizations can achieve a better culture. Culture was grown in suspension by a circular type, circular adherent growth type, as well as polygons adherent growth type composed of three different types of cells. By changing fluid cultures of ovarian cells in vitro longest survival of more than 1 year, and passaged three times. (4) Johnson dendrolimi-embryo and new hatched larvae cell culture: found by screening, trypsin digestion in of Johnson dendrolimi embryo tissue discrete processing, round suspended growth type, round adherent growth form, and fusiform adherent growth of 3 different types of cells. Embryo cells in vitro by changing the liquid culture has survived for more than eight months, and passaged twice; newly hatched larvae organizations use circular suspended growth type cells to move out from the tissue blocks the enzyme digestion process. Through the medium was changed culture found that newly hatched larvae cell proliferation in vitro is slow and difficult subcultured. In contrast, the pupal ovarian and embryo tissue cells easier to cultivate possible to obtain a continuous cell line proliferation. 2, the design and implementation of the insect cell database management system for the insect cell library routine maintenance work deficiencies in data and information management, using Microsoft Visual Basic 2005 software development tools and Microsoft SQL Server 2005 database management systems and other tools to build a an insect cell library management system, divided into two major sub-function system, 10 functional modules. (1) cell cryopreservation and recovery file management subsystem: four functional modules monitor the entire process of cell cryopreservation and recovery operation can achieve data add, modify, delete, search, statistics, and cell recovery operation timer to remind other functions. (2) the cell file management subsystems: six functional modules, each cell line data, including pictures, documents, information, as well as a variety of experimental data unified management and preservation, can add, modify, delete , export and other related operations. The system has a full-featured, simple to operate, convenient and practical features, and greatly improve the management level of the cell library, to lay the foundation for long-term effective management of insect cell library.
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