Construction of a Kunming Schizothorax fin cell line and ultra-low temperature cryopreservation method

Abstract

The invention relates to construction and ultralow temperature freezing preservation method of a fin cell line of schizothorax grahami, and belongs to the technical field of culture and ultralow temperature ultralow temperature freezing preservation of cells of fresh water aquatic organisms. The method comprises the following steps of: culturing in a DMEM(Dulbecco Modified Eagle Medium) / F12 culture solution which contains fetal calf serum and cell growth factors and has the pH value of 7.0-7.2 by taking the ventral fin tissue of the schizothorax grahami as a material and adopting a tissue explant method; carrying out subculture by adopting a trypsin digestion method. The method particularly comprises the steps of cell culture solution preparation, primary culture and subculture. The construction method disclosed by the invention is short in primary culture time consumption and large in cell quantity and can be used for chromosome analysis; the constructed fin cell line of the schizothorax grahami is in a fiber-like form, can be subjected to serial passage and directly applied to biological characteristic research, meets the requirements for germplasm resource conservation and theoretical research and application of the schizothorax grahami and is suitable for constructing the fin cell lines of other fishes.

Description

Technical field: [0001] The invention relates to the construction of a Kunming Schizothorax fin cell line 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 specie...

AI Technical Summary

Problems solved by technology

After years of hard work, Kunming Schizothorax artificial reproduction technology was successfully developed in 2013, and it is planned to release the breeding fry back to the Dianchi Lake and Niulan River and other watersheds. Abnormalities, such as polyploidy and aneuploidy, require short-term cell culture for karyotype analysis to test the quality of artificially propagated species, and ensure the survival rate and reproduction rate of released individuals; on the other hand, it takes a period of time for wild-introduced individuals to adapt Pond breeding environment, and this adaptation stage is the outbreak period of various diseases, which may bring disaster to the introduced individuals. How to prevent and control diseases is particularly important, and the research on the mechanism of disease resistance of drugs is an important part of disease prevention and control. It is indispensable to study the anti-disease mechanism of drugs at the molecular and cellular level. Therefore, it is necessary to study a method for the construction of a Kunming Schizothorax cell line and establish a Kunming Schizothorax cell line.

[0004] Through document retrieval, do not see identical report with the present invention