Research On The Culture Technology Of East Asian Migratory Locust Ovarian Cells Based On Microfluidic Chip

Insect cell culture technology is one of the important technical means in the field of life sciences,which has been widely used in the research of biology,pharmaceutical development,agriculture and animal husbandry and other fields.To meet the needs of certain applications and disciplines,it is still necessary to discover insect cells with specific insect species sources or specific functions.The cell culture technology based on the microfluidic chip is one of the cutting-edge cell culture technologies in cell biology research,and it has many unique advantages,such as high-precision microfluidic of fluid,automation,and low reagent consumption.The application of microfluidic chip to insect cell culture will help researchers expanding the research of insect cell culture and its biotechnology,and develop the huge potential in it.Locusta migratoria manilensis is one of the serious agricultural pests,and it is also a good experimental model.However,its cytological research is relatively lacking,and there is no report on the successful establishment of Locusta migratoria manilensis related cell lines.We selects locust to study culture of its ovarian cell in vitro using microfluidic technology.We designed a microfluidic chip that can be used in culture of animal cell,then develops culture of ovarian cell of Locusta based on microfluidic chip.Research on cell culture technology of migratory locust ovary.The main findings are as follows:1.Establishment of an in vitro culture method for ovarian cells of Locusta migratoria manilensis.Using the traditional culture platform,by studying the primary culture conditions of the ovary tissues of Locusta migratoria manilensis,a suitable culture method was obtained.The body surface of Locusta migratoria manilensis was disinfected with 75%ethanol for 50 minutes,and the ovarian tissues aseptically dissected were again disinfected with 75%ethanol for 5 minutes.The ovarian tissue is directly cut into tissue pieces and cultured in a culture bottle.The amount of medium added during inoculation is less than 0.5 ml,and the subsequent culture is gradually supplemented with medium.The medium is L-15 as the basic medium and 10%serum is added.The results showed that the ovarian cells of Locusta migratoria manilensis began to grow adherently after 3 days of culture,and cells of different morphologies could be observed to migrate from around the tissues at about 5 days.The tissues or cells adhered tightly and began to confluence with each other after 16 days of culture.2.Design,processing and testing of microfluidic chips.SolidWorks software was used to design the microfluidic chip and three-dimensional modeling,and the fluid dynamics simulation and processing technology optimization of the chip were performed.Finally,the human neuroblastoma cell line SH-SY5Y was used to evaluate the cell culture effect of the microfluidic chip.The results show that PMMA and PS are selected as the preparation materials of microfluidic chips,the thickness of the chip cavity is 8mm,and the chip processing is completed by thermocompression bonding at 180? for 20 minutes and epoxy resin potting glue.SH-SY5Y cells were successfully cultured on the microfluidic chip,and the cell morphology was normal,sustainable proliferation,and no microbial contamination problems occurred,indicating that the microfluidic chip designed in this paper can meet the needs of animal cell culture.3.Locusta migratoria manilensis ovary cell culture technology based on microfluidic chip.On the basis of the above-mentioned culture technology and chip design and processing,the research on the ovary cell culture technology of Locusta migratoria manilensis based on microfluidic chip has been carried out.The results showed that ovarian cells were cultured on the chip until about the 8th day,and tissue masses could be observed to adhere to the wall.Under suitable conditions,the current growth time of the ovarian cells of Locusta migratoria manilensis on the microfluidic chip could reach more than 30 days.It shows that the microfluidic chip has the potential to be widely used in a variety of insect cell culture,and provides a new research platform for insect cell culture.The method of in vitro culture of Locusta migratoria manilensis ovary cells established in this study can be applied to the cell biology research of Locusta migratoria manilensis in the future,and it also provides technical reference for the establishment of cell lines;the microfluidic chip designed and processed at the same time has a wide range of applications The potential for a variety of adherent cell cultures provides a new research platform for insect cell culture.