Tumor tissue 3D culture method and culture solution
A tumor tissue and culture method technology, applied in the field of biology, can solve the problems of easy floating of tissue samples, long period of species differences, poor tissue fixation, etc., to promote the process of research and development and precise treatment, easy operation, low cost effect
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Embodiment 1
[0043] A method for 3D culture of intestinal cancer tissue is provided, comprising the steps of:
[0044] (1) Preparation of tissue culture plate
[0045] Take a 24-well plate, add 5-10 μL of phosphate buffer solution dropwise, put the high-temperature sterilized circular cover glass in the well plate, pat it to fit the bottom, and set it aside for later use;
[0046] (2) Preparation of tumor tissue blocks
[0047] Take intestinal cancer tumor tissue, wash it with PBS containing 1% double antibody and gentamicin, and cut it into 0.5-1 mm with ophthalmic surgical instruments 3 The tissue pieces were washed with normal saline, and the whole operation was performed on crushed ice.
[0048] (3) Tumor tissue block culture
[0049] The following is a comparative experiment of screening gels, and a comparative experiment is conducted for three commonly used gels for cell culture: Matrigel, hydrogel, and vitro-Gel 3D-RGD:
[0050] 3.1 Matrigel glue
[0051] Add 30-100 μL of undil...
Embodiment 2
[0063] A method for culturing various tumor tissues in 3D is provided, comprising the steps of:
[0064] (1) Preparation of tissue culture plate
[0065] Take a 24-well plate, add 5-10 μL of phosphate buffer solution dropwise, put the high-temperature sterilized circular cover glass in the well plate, pat it to fit the bottom, and set it aside for later use;
[0066] (2) Preparation of tumor tissue blocks
[0067] Take intestinal cancer, gastric cancer and esophageal cancer tumor tissues, wash them with PBS containing 1% double antibody and gentamycin, and cut them into 0.5-1 mm with ophthalmic surgical instruments 3 The tissue pieces were washed with normal saline, and the whole operation was performed on crushed ice.
[0068] (3) Tumor tissue block culture
[0069] Add 20-100 μL of hydrogel (diluted with 20% sterile sucrose solution 1:1-1:5) dropwise on the glass slide, carefully move the tumor tissue pieces of colon cancer, gastric cancer and esophagus cancer into the hy...
Embodiment 3
[0076] Example 3 Feasibility Analysis of 3D Cultured Tumor Tissues for Efficacy Evaluation of Antitumor Drugs
[0077] In Example 2, during the culture of intestinal cancer tissue, different concentrations and types of chemotherapeutic drugs were added to the culture medium, and the tissues were taken out at different times after culture, fixed in the above-mentioned fixative solution, and stained by HE staining, Ki67 immunohistochemical staining or Corresponding other staining, observe the tissue structure and cell morphology under the microscope, compare the morphological differences with the samples without anti-tumor drugs, evaluate the drug efficacy, count the number of proliferating cells, and calculate the cell proliferation rate, see Table 3, analyze the cell Feasibility of proliferation ratio as an evaluation method and index of antineoplastic drug potency. See for details Figure 13 , Figure 14 and Figure 15 .
[0078] table 3
[0079]
Number of K...
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