Method for primary culture of tumor cells

A tumor cell and primary culture technology, applied in the field of cell biology, can solve the problems of high cell volume requirements, difficult removal of necrotic tissue, and inability to guarantee the success rate of culture, so as to improve the success rate, obtain cells freely and flexibly, and cultivate short cycle effect

Inactive Publication Date: 2014-06-18
NORTHWEST UNIVERSITY FOR NATIONALITIES
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AI Technical Summary

Problems solved by technology

[0003] The disadvantages of the existing technology are: (1) The types of malignant tumor tissues are complicated, and the success rate of in vitro primary culture cannot be guaranteed to be 100%
(3) It is difficult to remove non-tumor tissue and necrotic tissue, which can significantly affect the success rate of in vitro primary culture
(4) The requirement for the amount of cells is relatively high, too much or too little is not conducive to the success of primary culture
(5) Existing methods are easily affected by the culture system, especially the quality and quantity of calf serum, the type of medium, tumor cell type and pollution, etc. are the main factors affecting the primary culture of tumor cells in vitro

Method used

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  • Method for primary culture of tumor cells
  • Method for primary culture of tumor cells
  • Method for primary culture of tumor cells

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Embodiment Construction

[0023] The present invention will be described in detail below in conjunction with specific embodiments.

[0024] Example renal clear cell carcinoma primary culture

[0025] 1. Patients with clear cell renal cell carcinoma underwent unilateral total or partial resection, and the tumor samples were collected within half an hour after surgical resection. The collection method was to take a 50 mL plastic sterile centrifuge tube with a cover and contain 20 mL of RPMI1640 cell culture medium (pre-cooled in a 4°C refrigerator in advance), the culture medium contains 10% Hyclone peptide bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin. In a sterile environment, cut tumor samples from non-necrotic parts, with a volume of 1cm 3 The above is placed in the sterile centrifuge tube, and the centrifuge tube is transported on ice and brought back to the laboratory quickly.

[0026] 2. In the biological safety cabinet, transfer the tumor sample to a 100mm cell culture dish (Corn...

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Abstract

The invention discloses a method for primary culture of tumor cells and relates to the field of cell biology. According to the method disclosed by the invention, a tumor sample is prepared into a single-cell suspension liquid by use of an enzyme method or a mechanical method, and then tumor cell bladders are prepared by use of a serum-free suspension culture method, wherein alkaline fibroblast growth factors, epidermal growth factors, insulin and bovine serum albumin need to be added during a culturing process, then the growth factors are removed, and the culture is implemented in a general culture medium to obtain adherent cells capable of realizing continuous passage. The method disclosed by the invention is short in culture period and has the effect of increasing the success rate of the primary culture of tumors.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for primary culture of tumor cells. Background technique [0002] Primary culture, also called primary culture, refers to the first culture of tumor cells or tissues taken directly from the body. The most commonly used primary cultures are tissue block culture and dispersed cell culture. Tissue block culture is to directly transplant the shredded tissue block on the wall of the culture bottle, and then culture it after adding the culture medium. Dispersed cell culture is to disperse cells by mechanical or chemical methods in tissue blocks. For the isolation of the most viable free cells from clinical biopsy or resected tumor tissue, the classic method is to digest the intercellular complexes with proteolytic enzymes (such as trypsin and collagenase), or to use metal ion chelators (such as EDTA) to remove Ca which is dependent on cell adhesion to each other 2+ , and then s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
Inventor 宋雷
Owner NORTHWEST UNIVERSITY FOR NATIONALITIES
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