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Method for separating lung cancer cell from hydrothorax

A technology for lung cancer cells and pleural effusion, applied in the field of tumor cell biology, can solve the problems of many other cells and tissue fragments, little application significance, low tumor cell purity, etc., and achieves the effect of good effect and low cost.

Inactive Publication Date: 2012-09-19
SHANGHAI CHEST HOSPITAL
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  • Abstract
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Problems solved by technology

[0006] Although the direct centrifugation method used in the past is simple and easy, there are many other cells and tissue fragments, and the purity of tumor cells is low, so the practical application is of little significance

Method used

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  • Method for separating lung cancer cell from hydrothorax
  • Method for separating lung cancer cell from hydrothorax
  • Method for separating lung cancer cell from hydrothorax

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0033] Experimental steps:

[0034] 1. Preparation of Percoll solutions with different concentrations (density):

[0035] First mix 9 parts of Percoll with 1 part of 8.5% NaCl or 1.5M PBS to achieve physiological osmotic pressure, and then dilute to the required concentration with physiological solution (0.85% NaCl or 0.15M PBS). We use 60% and 40% concentrations. The concentration gradient is found to be simple and effective through comparison, and the separation effect is better.

[0036] 2. Preparation of pleural fluid suspension:

[0037] Centrifuge the pleural fluid sample at 1500rpm for 5 minutes, discard the supernatant, wash with 1% PBS, centrifuge at 1500rpm for 5 minutes, discard the supernatant, and resuspend in 1% PBS; at this time, the pleural fluid suspension contains various cell components and cell tissue fragments.

[0038] 3. Preparation of discontinuous density gradient Percoll layer:

[0039] 60%, 40%, and pleural fluid suspensions were arranged layer b...

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Abstract

The invention discloses a method for separating lung cancer cell from hydrothorax. The method comprises the following steps of: (1) preparing Percoll solutions with different concentrations; (2) preparing hydrothorax suspension; (3) preparing Percoll layers with discontinuous density gradients; (4) centrifuging; (5) sampling; and (6) performing cell culture. The method disclosed by the invention is simple and stable and has the advantages of low cost and good effect.

Description

technical field [0001] The invention relates to a method for separating tumor cells, in particular to a method for separating and purifying lung cancer cells in pleural effusion, and belongs to the technical field of tumor cell biology. Background technique [0002] The thorax is a closed cavity composed of parietal pleura and visceral pleura, inside which is negative pressure. Under normal circumstances, there is a small amount of fluid (about 1-30 ml) between the two layers of pleura for lubrication , to reduce the friction between the two layers of pleura during the breathing process, which is conducive to the expansion and contraction of the lungs in the chest cavity. This fluid is produced from the parietal pleura and absorbed by the visceral pleura. It is in a dynamic balance after continuous circulation, and the amount of fluid remains constant. When something happens to affect the pleura, whether the parietal pleura produces pleural fluid or the rate at which the vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
Inventor 顾爱琴刘蕾李榕孙磊姚明
Owner SHANGHAI CHEST HOSPITAL
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