A CELL PRESERVATION SOLUTION AND THE FREEZING AND PRESERVATION PROCEDURES OF ANIMAL CELLS USING THE SAID SOLUTION

摘要

PURPOSE: A cell preservation solution and a method for freezing and preserving animal cells using the same are provided. The cell preservation solution consists of compounds of which molecular formula and weight are well known. Therefore, it is capable of storing cells stably for a long time. CONSTITUTION: A cell preservation solution comprises 3 to 10 mM of KH2PO4, 20 to 35 ml of KOH, 20 to 250 mM of NaCl, 0.4 to 4 mM of MgCl2, 15 to 35 mM of lactic acid, 3 to 15 mM of dextrose and 150 to 500 mM of sorbitol, and has pH 6.3 to 7.5. The cell preservation solution may further comprises 0 to 20 volume% of DMSO(dimethyl sulfoxide). A method for freezing and preserving animal cells comprises the steps of: (1) removing a cell-cultured medium from a cell culture dish; (2) washing the dish with a trypsin-EDTA solution to separate the cells from the dish; (3) suspending the separated cells in a fresh medium, transferring the medium into a vessel, and centrifuging it; (4) removing the supernatant; (5) suspending the cells in the cell preservation solution; (6) placing the suspended solution in ice for 10 minutes; (7) adding 0 to 20 volume% of DMSO into the suspended solution; (8) inserting the suspended solution into a vessel and freezing it at -80 deg. C; and (9) storing the frozen cell at -80 deg. C or in liquid nitrogen.