Separation method for mantle epithelial cells in vitro of pinctada martensii

Abstract

The invention relates to a separation method for mantle epithelial cells in vitro of pinctada martensii, belonging to the technical field of separation for histiocyte culture. The method mainly comprises steps of: (1) preparing Percoll gradient solution as follows: mixing nine parts of Percoll stock solution with one part of D-MBSS to obtain 100% of Percoll solution; and mixing the 100% Percoll stock solution with D-MBSS solution in different proportion to obtain the Percoll gradient solution; (2) preparing gradient layer as follows: previously using BSA solution to soak the tube wall of a centrifuge tube and preparing a gradient tube; (3) adding cell suspension as follows: adding cell suspension to be separated into the gradient tube; and (4) cell separation: after centrifugating, taking out cells, washing and heavy suspending. The invention can separate out single type of cell group from cells emigrated from primarily cultured tissue blocks, has low damage to cells, and can maintain activity of cells to the maximum extent. The cells after being separated still can be cultured. The operation is simple.

Description

technical field [0001] The invention relates to a tissue cell culture and separation technology, in particular to a technology for separating a single type of cell from a primary cultured tissue block of a pearl oyster, Hepu pinnacle oyster. Background technique [0002] Pinctada fucata Martenssi, also known as Pinctada fucata Martenssi, belongs to the phylum Mollusca, the class Bivalvia, the order Pterioida, the family Pteriidae, and the genus Pinctada ( Pinctada), mainly distributed in the coastal areas of my country, is currently the main shellfish used in the production of seawater pearls in the world. In recent years, due to the impact of global climate change and the economic purpose of cultivating shellfish, the quality of pearl oysters and the quality of pearls have declined. The culture and isolation of Hepu pearl oyster tissue cells are of great significance to the quality of pearl oyster and pearl. [0003] Because many cells migrated out during the culture proce...

AI Technical Summary

Problems solved by technology

[0003] Due to the large number of cells migrated out during the culture of Hepu pinnacle primary cells, it is not conducive to the isolation and culture of cells and the analysis of the properties of a single cell population

At present, there are very few studies on the separation and purification of Hepu pinnacle tissue cells, and the related technology is relatively weak