Method and its reagent unit for separating and purifying tumor cell
A technology for separation and purification of tumor cells, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve problems such as time-consuming, cross-contamination, and reduced applicability, and achieve high cell purity, high survival rate, and compatibility good sex effect
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Embodiment 1
[0044] Separation and purification of tumor cells from pathological tissues
[0045] 1. Digestion of tumor pathological tissue:
[0046] Surgical resected tumor pathological tissue (about 1 gram) was placed in a serum-free culture medium culture bottle. Visible non-tumor tissue and tumor necrosis tissue were removed with sterile scissors. Wet the tumor tissue with 10ml of serum-free medium and cut it into pieces as soon as possible. These shredded tumor tissues were placed in a suitable container, tumor tissue digester was added in an amount of 10 ml, and incubated in a 37° C. water bath for 4 hours. Then take out the container, dilute it twice with serum-free culture medium, and after repeated pipetting, filter the cell suspension through a sterile 100 μm cell filter, collect the cell filtrate, centrifuge at 1200 rpm for 8 minutes, pour Remove the supernatant, suspend these precipitated cells (which contain lymphocytes, fibroblasts, stromal cells and tumor necrotic cells) ...
Embodiment 2
[0052] Isolation and Purification of Tumor Cells from Malignant Ascites
[0053] 1. Preparation of Ascites Tumor Cell Suspension:
[0054] Take 20ml of malignant ascites obtained from the patient, put it in a suitable container, centrifuge at 1200 rpm for 8 minutes, pour off the supernatant, and take 5ml of tumor cell suspension to suspend these precipitated cells (which contain lymph cells, fibroblasts, stromal cells and tumor necrotic cells), and gently mixed to obtain tumor tissue cell suspension.
[0055] 2. Separation and purification of tumor cells:
[0056] Take a container containing 5ml of tumor cell separation agent, gently add 5ml of tumor tissue cell suspension from the tube wall to the top of the separation agent in the container, let it stand for 6 minutes, and suck out 1.5ml of the solution close to the bottom of the container, Place in another container and centrifuge at 1200 rpm for 8 minutes. The precipitated cells are the purified tumor cells.
[0057] 3...
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