Cell culture container and primary cell culture method
A cell culture and container technology, applied in the field of cell culture, can solve the problem of low yield of primary cells
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Embodiment 1
[0057] The structure of the cell culture vessel used for primary culture in embodiment 1 is as follows: figure 1 shown. The culture unit of the cell culture container in embodiment 1 is one, and the material of culture tank is polystyrene, and the diameter of culture tank is 100mm, and the depth of culture tank is 20mm, and the length of fixture is 10mm, and the diameter of fixture is 1mm , the diameter of the cover plate is 92mm, the diameter of the vent hole is 0.6mm, and the diameter of the fixing hole is 1.1mm.
[0058] The steps of the primary culture of embodiment 1 are as follows:
[0059] (1) Umbilical cord collection: collect full-term healthy cesarean section fetal umbilical cord (1.5 cm in diameter), immerse in PBS containing 1% (m / v) penicillin and 1% (m / v) streptomycin, place on ice.
[0060] (2) Tissue separation: cut the umbilical cord into 4 cm long sections in a clean bench, cut them longitudinally, rinse them repeatedly with sterile PBS until the liquid is...
Embodiment 2
[0069] The primary culture steps of Example 2 are roughly the same as those of Example 1, the difference being that the tissue used in Example 2 is amnion tissue during the culture process.
[0070] According to statistics, in Example 2, the amount of cells crawling out of every 4cm of umbilical cord tissue is 6.1×10 7 The adherence rate of the tissue block was 79.9%.
[0071] Table 2 shows the results of the flow cytometric identification of the third-generation mesenchymal stem cells in Example 2.
[0072] Table 2
[0073]
[0074]
[0075] As can be seen from Table 2, the positive rate of the surface expression of CD105, CD73, CD90, CD45, CD34, CD14, CD19 and HLA-DR in the third generation mesenchymal stem cells of Example 2 is in line with international standards (specified in international standards, mesenchymal The cell surface of stromal stem cells expresses CD105, CD73 and CD90 (≥95%), and does not express CD45, CD34, CD14 or CD11b, CD79α or (≤2%)).
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