Cell strain for virus isolated culture and propagation
A virus isolation and virus culture technology, applied in the field of animal husbandry, can solve the problems of complicated and backward manufacturing process, high production cost, and carrying other pathogens, and achieve stable cell shape and growth speed, long growth cycle and good growth performance Effect
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Embodiment 1
[0029] Cloning and selection of Vero cell lines
[0030] 1. Cloning culture of Vero cells by limiting dilution method
[0031] The Vero cells preserved in the laboratory without mycoplasma, bacteria, fungi, and exogenous viruses were digested with trypsin to make a single-cell suspension. After counting the cells, they were serially diluted, and then inoculated in 96-well cell culture plates at 37°C. 5%CO 2 After culturing in an incubator for 15 days, the cells in the cell wells with single cell growth were selected and transferred to a 24-well plate for culture, and the cells were expanded and cultured sequentially to obtain subcloned cell lines.
Embodiment 2
[0033] Study on Biological Properties of African Green Monkey Kidney Cell Line VeroB10
[0034] 1. cell morphology
[0035] The African green monkey kidney cell line VeroB10 strain grown into a monolayer was prepared into a cell suspension, added to DMEM growth medium containing 8% fetal bovine serum, and incubated at 37°C in 5% CO 2 Under the condition of culture for 48h, observe the cell morphology. The cells are well adherent, plump in shape, clear in boundary and smooth in surface. Compared with Vero mother cells, VeroB10 strain had more obvious and orderly boundaries, strong refraction, full cytoplasm and clear nucleoplasm.
[0036] 2. Cell Kinetic Detection
[0037] Make a cell suspension from VeroB10 and parental Vero cells covered with a single layer. After counting the cells, count the cells as 5×10 4 / mL was inoculated into a 24-well plate, added to DMEM medium containing 8% newborn calf serum, and incubated at 37°C in 5% CO 2 Cultivate under conditions, take ...
Embodiment 3
[0057] Lesion Characteristics of Goat Pox Virus in VeroB10 Kidney Cells of African Green Monkeys
[0058] The African green monkey kidney cell line VeroB10 was inoculated in a 6-well plate, added to DMEM cell growth medium containing 8% fetal bovine serum, and incubated at 37°C in 5% CO 2 After 24 hours under the same conditions, it can grow into a single layer. Inoculate 0.1mL goat pox virus AV41 strain, add DMEM maintenance solution containing 2% fetal bovine serum, and culture at 37°C for 7 days to observe the cytopathic changes every day. Lesions appear, and the main lesion feature is the shrinkage of cells into piles.
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