Organoid culture system and organoid culture method
A culture system and organ culture technology, applied in the field of organoid and cell culture, can solve the problems of unfavorable large-scale culture and application of organoids, complicated culture steps, high culture cost, etc., and achieve simplified culture process and culture cost, easy conditions, Cultivate effects with simple steps
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Embodiment 1
[0113] Example 1: 3-D Culture of Kidney Tissue Organoids
[0114] Step 1, 293T cells were revived, cultured with IMDM (10% (v / v) fetal bovine serum), and passaged.
[0115] Step 2: Select 293T cells in a well-cultured and well-grown culture dish, wash them twice with PBS, and digest with 0.25% trypsin. Stop the reaction.
[0116] Step 3, collect the cell suspension into a centrifuge tube, centrifuge at 1000rpm for 5min, discard the supernatant, wash the cells with PBS, then centrifuge at 1000rpm for 5min, repeat washing twice, and then resuspend with 1ml medium cells, and count the cells using a cell counter.
[0117] Step 4, according to the number of cells, dilute the cells with medium to 100 μl system containing 80 cells to obtain a cell suspension for later use.
[0118] Step 5, select Matrigel ( Growth Factor Reduced (GFR) Basement Membrane Matrix, Phenol Red-free, LDEV-free, Life Sciences; 356231) was used as Matrigel, first take 32 μl Matrigel, spread it in a 96-...
Embodiment 2
[0120] Example 2: 3-D Culture of Kidney Tissue Organoids
[0121]The difference between this example and Example 1 is that the matrigel used in step 5 is RGF-BME (CultrexReduced Growth Factor Basement Membrane Extract, Type 2, Pathclear R&D; 3533-005-02).
Embodiment 3
[0122] Example 3: 3-D Culture of Kidney Tissue Organoids
[0123] The difference between this example and Example 2 is that in step 4, the cells are diluted to a 100 μl system containing 800 cells.
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