Method for establishing schizothorax wangchiachii heart cell line and performing ultralow temperature cryopreservation on schizothorax wangchiachii heart cell line

Abstract

The invention relates to a method for establishing a schizothorax wangchiachii heart cell line and performing ultralow temperature cryoprservation on the schizothorax wangchiachii heart cell line, belonging to the technical field of freshwater aquatic organism cell culture and ultralow temperature cryopreservation. The method comprises the following steps of taking heart tissues of schizothorax wangchiachii as materials, and culturing the materials in an M199 culture solution by adopting a tissue block method and a two-enzyme digestion method, wherein the M199 culture solution contains fetal calf serum and cell growth factors and the pH value of the M199 culture solution is 7.0-7.2; and performing secondary culture by adopting a trypsase digestion method. The method specifically comprises the steps of preparing a cell culture solution, performing primary culture and performing secondary culture. The method has the beneficial effects as follows: 1, the operation is simple and convenient; 2, damages caused by a pure tissue block method to heart cells are reduced; 3, the form of the established schizothorax wangchiachii heart cell line is in the shape of fibroblasts, and the established schizothorax wangchiachii heart cell line grows well, can realize continuous passage culture, can be directly applied to biological characteristic researches, and meets requirements for storage, theoretical study and application of genetic resources of the schizothorax wangchiachii; and 4, the establishing method is also suitable for establishing heart cell lines of other types of fishes.

Description

Technical field: [0001] The invention relates to the construction of a heart cell line of Schizothorax brevista and an ultra-low temperature cryopreservation method, belonging to the technical field of freshwater aquatic organism cell culture and ultra-low temperature cryopreservation. Background technique: [0002] Fish cell culture started in the 1960s, and has developed a relatively complete cell culture system including culture medium, antibiotics, primary culture and subculture methods. Up to now, more than 280 cell lines have been established. After more than 30 years of fish cell culture in my country, only more than 50 fish cell lines have been established, involving about 30 species, mainly marine species. From the above examples, it can be seen that a mature cell culture process is not a prerequisite for the successful establishment of a cell line. A mature technology process cannot make cell culture successful, and cell culture still needs to provide primary cell ...

AI Technical Summary

Problems solved by technology

Schizothorax is a bottom-dwelling fish. It generally lives in canyon rivers with relatively fast currents. It is often found in deep pools impacted by rapids in tributaries. There is still a certain amount in the distribution area, but its production has been greatly reduced

In 2007, the Yunnan Fisheries Research Institute succeeded in artificial repro