Novel three-dimensional culture method of glioma stem cells and application thereof
A technology of glioma stem cells and glioma cells, which is applied in the field of cell biology, can solve the problems of difficult observation of cell spheroids and poor repeatability of spheroid culture, so as to reduce the waste of patient resources and follow-up research funds, and achieve good biocompatibility sexual effect
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[0025] Example 1
[0026] A novel three-dimensional culture method for glioma stem cells uses a three-dimensional collagen scaffold and finds a suitable low serum culture concentration. The culture method includes the following steps:
[0027] 1) Cut the three-dimensional collagen scaffold into 1*5*5mm cubes, irradiate and disinfect with Co60 overnight before use, and store in a dry environment at 4°C for later use. Before inoculating cells, pre-process with cell culture solution for 12 hours;
[0028] 2) Digest and centrifuge the glioma cell line to a single cell suspension, and then wash it twice with PBS to remove serum components, and perform cell counts. Each three-dimensional collagen scaffold is 1×10 4 ~5×10 4 Inoculate the glioma cells on the three-dimensional collagen scaffold treated in step 1) and let it stand for 4 hours. After the cells have effectively adhered to the scaffold, they are placed in a cell culture plate for culture and replaced every 2 days. After sub-cultu...
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[0033] Example 2
[0034] This example is an optimized solution based on Example 1, and provides a new three-dimensional culture method for glioma stem cells. For the same parts in this embodiment as Embodiment 1, please refer to the public content in Embodiment 1 for understanding. The public content in Embodiment 1 should also be the content of this embodiment, and the description will not be repeated here.
[0035] The three-dimensional culture method includes the following steps:
[0036] The conventional two-dimensional cultured U87 cells were digested and centrifuged to prepare a single cell suspension, and then seeded on a collagen scaffold with a pore size of about 50 μm, and cultured under the condition of 1% fetal bovine serum (FBS) + DMEM medium. Observe whether the cells can adhere to the scaffold collagen matrix and grow well, observe the proliferation of U87 cells transferred into green fluorescent protein (GFP) by fluorescence microscope, observe the cell growth morph...
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