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Primary cell isolation and medium this step
Primary cell isolation and medium this step
1, the choice of organs and tissues
Remove unnecessary tissue and blood as much as possible.
2. Separation of primary cells
(1) The PriCells primary cell isolation kits I, II, and III were used.
(2) Different PriCells primary cell isolation kits may be used depending on the tissue source.
3. Cultivation of primary cells:
(1) PriCells primary cell special medium
(2) PriCells primary cell special additives
(3) high quality fetal bovine serum
4. Cell culture and identification:
PriCells Primary Cell Identification Kit
4. Passage and preservation of primary cells
(1) Passage: According to the growth state of the cells, the grown cells were passaged 1:2 or 1:3.
(2) Preservation: DMSO + culture solution + serum
Cool down in the following order: room temperature → 4 ° C (20 minutes) → refrigerator freezer (30 minutes) → ultra-low temperature refrigerator (-80 ° C overnight) → liquid nitrogen.
5. Recovery of primary cells
(1) Remove the cells and quickly thaw them in warm water at 37 °C.
(2) Aspirate the cell suspension and add 10 times or more of the culture solution.
(3) After appropriately diluting with the culture solution, the culture flask was inoculated and placed in a 37 ° C CO 2 incubator for static culture.
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