Human primary tumor cell separation and culture kit

A primary tumor cell, cell culture technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve problems such as cell contamination, achieve high purity, simple and convenient culture effect, and overcome the effect of complex culture process

Inactive Publication Date: 2012-07-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the above process, a variety of consumables and reagents are needed, including cell culture dishes or culture plates, diges...

Method used

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  • Human primary tumor cell separation and culture kit
  • Human primary tumor cell separation and culture kit
  • Human primary tumor cell separation and culture kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] see figure 1 , figure 2 , The kit consists of packing box 1, 1 piece of 6-well cell culture plate 2, 5ml cleaning solution 3, 1 bottle of 5ml trypsin digestion solution 4, 1 bottle of 5ml collagenase-hyaluronidase digestion solution 5, 1 bottle of 5ml cell separation Solution 6, 1 bottle of 5ml tumor selective culture solution 7, instruction manual 8.

[0020] The cleaning solution 3 is a sterile RPMI 1640 cell culture solution containing 400 U / mL of penicillin and 400 ug / mL of streptomycin, filled into two 5ml sterile sealed bottles, labeled, and stored at 4°C.

[0021] Trypsin digestion solution 4 is prepared with sterile ultrapure water containing 0.1% trypsin (purchased from Sigma, 100mg), 10g / L polyvinylpyrrolidone trypsin digestion solution, divided into 5ml sterile sealed bottles, and affixed Label and store at 4°C.

[0022] Collagenase-hyaluronidase digestion solution 5 is prepared with RPMI 1640 cell culture medium, wherein the final concentration of collag...

Embodiment 2

[0027] see figure 2 , using this kit: first number each well of the 6-well cell culture plate 2 (a-f).

[0028] (1) After cutting out the tumor specimen, put it into well a of the 6-well cell culture plate 2, and pour cleaning solution 3 to cover the specimen;

[0029] (2) Wash the tumor specimen in well 1, move the specimen into well b, pour cleaning solution 3, and chop or cut the tumor specimen into 1mm 3 size;

[0030] (3) Move the cleaned and shredded tumor specimen block into well c, pour trypsin digestion solution 4, and pre-digest in a cell culture incubator at 37°C for 15 minutes;

[0031](4) After removing from the cell incubator, transfer the pre-digested tumor specimen into well d, pour collagenase-hyaluronidase digestion solution 5, and digest overnight in a refrigerator at 4°C;

[0032] (5) After overnight digestion, collect the digested cell suspension, filter it through a 200-mesh cell strainer into a 50ml centrifuge tube, add cell separation solution 6 to ...

Embodiment 3

[0036] Liver cancer tissue specimens were taken, and the operation steps were the same as those in Example 2 to obtain high-purity liver cancer cells growing in suspension. See results Figure 4 , is the primary liver cancer cells cultured in well f, in the figure: C is the suspended liver cancer cells.

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Abstract

The invention provides a human primary tumor cell separation and culture kit, consisting of a packing box, a cell culture plate with 6 pores, cleaning solution, trypsinization solution, collagenase-hyaluronidase digestive juice, a cell separating medium, selective tumor culture solution and an operating instruction, wherein sterile RPMI (Roswell Park Memorial Institute) 1640 cell culture solutioncontaining 400U/mL of penicillin and 400 mu g/mL of streptomycin serves as the cleaning solution. The human primary tumor cell separation and culture kit solves the problems that the conventional primary tumor cell culture process is complicated, the purity of obtained tumor cells are not high, and the tumor cells are easy to be contaminated by cells. According to the invention, the design is reasonable, the opportunity of germ contamination is less in the process of separation and culture of primary tumor cells in vitro, and only a few of centrifugal tubes and other common consumable items are needed, so that the kit is simple and convenient to use, the purity of the cultured tumor cells is higher, and the effect is good.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a kit for separating and culturing human primary tumor cells. Background technique [0002] Tumor cell culture is an important means to study the mechanism of carcinogenesis and the biological characteristics of tumor cells. The application of in vitro culture technology for tumor research has many advantages: (1) It can be free from the influence of internal factors in the body, so that it is convenient to explore various fields such as physics, chemistry and biology. (2) It is convenient to study the structure and function of tumor cells; (3) It can be stored for a long time to observe the changes of tumor cell genetic behavior; (4) It can be used for rapid screening of anticancer drugs. [0003] Tumor cell culture technology is divided into primary tumor cell culture and tumor cell line culture. Primary tumor cell culture refers to directly separating tumor cells from livin...

Claims

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Application Information

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IPC IPC(8): C12N5/09
Inventor 丁国平曹利平
Owner ZHEJIANG UNIV
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