Method for constructing cell line by using insect egg

Abstract

The invention relates to a method for establishing a cell line of insect eggs, which comprises the following steps: 1) dipping an insect oosperm sheet which has been laid for 90 to 100 hours in a sodium hypochlorite solution or a formaldehyde solution, and then disinfecting and washing the oosperm sheet; 2) pouring the treated eggs into an insect cell culture solution, and extruding the eggs one by one to release embryos; 3) cutting the embryos into tissue blocks to be cultured in an incubator; 4) adding the cell culture solution into the incubator to culture the tissue blocks continuously; 5) replacing the culture solution periodically until the proliferating cells are full of the whole culture bottle; 6) sucking out half of the cell culture solution containing newly proliferated single cells, putting the cell culture solution into a new culture bottle which is added with the same amount of a new cell culture solution, and putting the new culture bottle back to the incubator to culture the cells continuously; and 7) repeating the step 5) and the step 6) periodically to cause the cells to begin to passage so as to establish the cell line successfully, wherein the whole process is performed under a sterile condition. The insect eggs can be lepidoptera insect eggs. The method shortens the time from the prior 1.5 to 2 years to 1 to 3 months, thereby having quite obvious superiority and strong repeatability.

Description

technical field [0001] The invention relates to a biotechnology, in particular to a method for establishing cell lines with insect eggs. Background technique [0002] Since the first insect cell line was successfully established in 1965, more than 500 insect cell lines have been established in the past 40 years. As a research material, insect cell lines have always been an important tool for scientific research in physiology, developmental biology, cell biology, molecular biology and biochemistry, and insect cells, as an important part of the baculovirus expression vector system, express a large number of exogenous proteins of great economic or scientific significance; at the same time as a bioreactor to amplify insect baculoviruses for use as biopesticides, especially to amplify recombinant baculovirus insecticides containing exogenous genes, insects Cells also play an important role. [0003] A large number of commercially available cell lines from Lepidoptera insects ha...

AI Technical Summary

Problems solved by technology

Therefore, whether the establishment of cell lines is successful or not is very random and has great uncertainty. In most cases, it ends in failure, and the process is very long. Generally, it takes 1.5-2 years or even longer for a cell line to be successfully established.