Rapid purification method of high-purity primary tumor cells

Abstract

The invention relates to a rapid purification method of high-purity primary tumor cells. The method comprises the following steps of oscillating and digesting tumor tissues to prepare a mixed cell suspension; slowly adding the mixed cell suspension to the upper layer of fetal calf serum along the wall of a centrifuge tube; after addition, putting the mixture in an incubator for still standing; andperforming time difference digestion on adherent cells to obtain the high-purity primary tumor cells. According to the rapid purification method of the high-purity primary tumor cells provided by theinvention, an oscillator is used for digesting tissue masses, so that the tissue masses can be accelerated to be dispersed into the single cell suspension, the damage of mixed digestive enzymes to cell membranes is reduced, the survival rate of inoculated cells is improved, and the characteristics of primary cell membranes can be protected; and the purification of the primary tumor cells is realized by fetal calf blood serum still standing precipitation and time difference pancreatin digestion, the tumor cells and other non-tumor cells can be separated to the maximum degree, the purificationefficiency is high, and the purification time is saved. The method is simple and practical, and provides important technical support for application to individualized treatment of clinical patients.

Description

technical field [0001] The invention belongs to the technical field of purification of primary tumor cells, and in particular relates to a rapid purification method of high-purity primary tumor cells. Background technique [0002] In today's society, cancer is one of the important diseases that threaten human health, and its morbidity and mortality remain high. Since the specific characterization of tumor cells is difficult to find, a specific therapeutic drug for cancer has not been found in the existing scientific research. [0003] The primary tumor cell culture method refers to the method of extracting tissues from animals under sterile conditions, simulating the physiological environment in vivo to make them survive and grow, and maintain their structure and function. As an important means of cancer research in vitro, it can It provides an effective cell source for in-depth research and treatment of tumors, and complements in vivo research. If the primary tumor cells ...

AI Technical Summary

Problems solved by technology

However, the success rate of the existing primary culture of tumor cells is not high, and it takes a long time. When the tumor cells grow stably, they have lost many of the unique biological characteristics of the tumor tissue. Therefore, we still need to further explore the relationship between Study how to shorten the purification and culture time of primary tumor cells to ensure that their shape is similar to that in vivo, and provide an effective platform for the study of tumor occurrence, development and metastasis mechanisms, detection of anti-tumor drug sensitivity, and even tumor gene therapy

[0005] Although the technical methods for establishing tumor cell lines are constantly improving, there is still no efficient and standardized establishment method. The existing cell purification methods are mainly divided into two types: natural purification and artificial purification.

As the earliest purification method, natural purification takes advantage of the proliferation advantages of a certain type of cells to naturally eliminate inferior cells in the long-term passage process, leaving only vigorously growing cells to achieve cell purification, but this method takes a long time and remains Almost all of them are fibroblasts, only malignant tumor cells or mutated cells can be preserved by this method, and continuous purification is required to meet the standards for establishing cell lines. Excessive culture time will affect the cytology of primary tumor cells properties, studies that affect their biological properties

Artificial purification uses artificial technology to create a favorable living environment for the growth of a certain type of cells, and realizes the purification of cells. At present, the following methods are mainly used: enzymatic digestion, mechanical scraping and cloning, etc., but all have the following shortcomings: Enzyme digestion usually uses trypsin, which acts on the peptide bond composed of arginine and lysine, and has a very strong digestion ability, but it does a lot of damage to the cell membrane, which greatly affects the research on the biological characteristics of the tumor cell membrane surface; The scraping method is to directly scrape off the fibroblast clusters and other cell clusters other than the tumor cell clusters under the microscope, but this method has high requirements for the operator's work experience, and requires the operator to have in-depth research on the growth of cells , and can judge different cell groups through experience, and the human error is relatively large; the cloning method divides the mixed cells into several cell suspensions, and obtains the tumor cell clone group required for the experiment through the inspection of different clone cell groups, and the cells obtained by this method The clone is very pure, but it takes a long time. During the long-term culture process, the biological characteristics of tumor cells are significantly different from those just isolated from the body, which affects the accuracy of the research.

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