A method for constructing a kidney cell line of Schizothorax dali

Abstract

The invention relates to a construction method of a kidney cell line of Schizothorax taliensis. The method comprises following steps: 1), obtaining of kidney tissue of the Schizothorax taliensis: potassium permanganate and alcohol are combined to disinfect the Schizothorax taliensis; 2), primary culture: an L-15 or DMEM / F12 culture solution containing cell growth factors, penicillin, streptomycin and amphotericin B is adopted for culture, and osmotic pressure of the culture solution is 280-300 mOsm / L; 3), subculture: when subculture is performed to the tenth generation, the cell culture fluid is replaced with a basic culture solution, and the kidney cell line of the Schizothorax taliensis is constructed successfully. With the adoption of the construction method, the form of the obtained kidney cell line of the Schizothorax taliensis is fibroid, the kidney cell line can realize continuous passage, can be directly applied to research of biological characteristics, can meet the requirements for germplasm resource preservation and theoretical research of the Schizothorax taliensis as a rare species and is the first cell line of the Schizothorax taliensis, thereby laying the foundation for research of plateau adaptability of the cell level of the Schizothorax taliensis.

Description

technical field [0001] The invention relates to a method for constructing a kidney cell line of Schizothorax dali, belonging to the technical field of freshwater aquatic organism cell culture and ultra-low temperature cryopreservation. Background technique [0002] Dali Schizothorax (Schizothorax taliensis) belongs to Cyprinidae Schizothorax subfamily Schizothorax, originally a special economic fish in Erhai Lake, accounting for 30% of Erhai Lake's catches in history. In the 1970s, due to the drop in the level of the Erhai Sea, dozens of ditches and fish holes dried up, the Dali schizothorax lost its spawning grounds, and the introduction of foreign species devoured a large number of Dali schizothoracidae eggs. The number of Dali Schizothorax has dropped sharply. According to the collection records of the Fish Specimen Bank of Kunming Institute of Zoology, Chinese Academy of Sciences, since the last collection in 1973, no Schizothorax Dali has been found in 43 years. It is ...

AI Technical Summary

Problems solved by technology

It can be seen that the construction speed of cell lines is very slow, and insufficient attention and fewer workers may be one of the reasons; secondly, compared with mammalian cell culture, fish cell culture is more prone to contamination, This leads to an increase in the failure rate, and most of the existing patents or articles attribute this phenomenon to poor technology, and pay less attention to the quality of the fish body itself. In the practice of Dali Schizothorax cell culture, the living conditions of the fish body It also directly affects the success or failure of the cell line construction; in addition, due to the limitation of the wild habitat of Dali Schizothorax, little is known about its life habits, which also increases the difficulty of Dali Schizothorax cell culture to a certain extent

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