Method for separating salivary gland cells of hirudo nipponia

Abstract

The invention discloses a method for separating salivary gland cells of hirudo nipponia, and relates to the technical field of cell separation. The method comprises the following steps: S1, putting extracted salivary gland tissue into a pre-cooled buffer solution; S2, washing off bloodstains of the salivary gland tissue by using the buffer solution; S3, transferring the salivary gland tissue treated in the step S2 into a culture dish; S4, adding a disinfectant for disinfection; S5, performing cleaning again by using the buffer solution, and washing off the residual disinfectant; S6, transferring the salivary gland tissue treated in S5 into a centrifuge tube, adding a proper quantity of an enzyme solution, and performing water bath enzymolysis; S7, taking supernatant treated in the S6, andadding a culture solution at the same time; and S8, performing centrifugal treatment on the salivary gland tissue treated in the S7, discarding the supernatant, adding a proper quantity of a culture solution, performing resuspending, and performing inoculating to a culture plate. According to the method for separating the salivary gland cells of the hirudo nipponia, provided by the invention, thesalivary gland cells are further cultured, active substances such as hirudin and the like can be extracted, and the method can be used for researching a salivary gland cell secretion mechanism.

Description

technical field [0001] The invention relates to the technical field of cell separation, more specifically, it relates to a method for isolating salivary gland cells of Hirudo japonica. Background technique [0002] With the intensification of population aging and the change of living environment, the incidence of cardiovascular and cerebrovascular diseases is also showing an increasing trend year by year, such as hypertension, cerebral thrombosis, cerebral hemorrhage, etc. There are more and more patients, and the corresponding prevention and treatment measures to solve the disease Urgently important. Studies have shown that hirudin is the strongest anticoagulant active substance found in the world so far, and it is the most effective drug raw material for treating cardiovascular and cerebrovascular diseases. Japanese medical leeches are widely distributed in China and are traditional Chinese medicines in my country. With their strong anti-platelet aggregation and antithrom...

AI Technical Summary

Problems solved by technology

The main medicinal part of Japanese leeches lies in its salivary glands, but because the research on Japanese leeches is very scarce at this stage, and mainly focuses on the artificial breeding of Japanese leeches and the research on biologically active components, the salivary glands of Japanese leeches There are few reports on the isolation and cultivation of relevant research, which is not conducive to the development and utilization of Japanese medical leech resources