Cell preserving fluid for delaying cell growth in cell culture and preparation method thereof
A technology of cell culture and preservation method, which is applied in the field of cell preservation solution for delaying cell growth and its preparation, which can solve the problems of excessive cell growth and achieve the effects of maintaining activity, reducing the cost of cryopreservation, and prolonging the time
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[0038] The present invention also provides a method for preparing a cell preservation solution, the steps of the preparation method comprising:
[0039] Take sodium thiosulfate, mannitol, sucrose, human serum albumin, and phosphate buffer in specified proportions, and then dissolve sodium thiosulfate, mannitol, sucrose, and human serum albumin in phosphate buffer to obtain cell preservation liquid;
[0040] Store the cell preservation solution at less than or equal to 5°C, sealed and protected from light.
[0041] The present invention also provides a cell preservation method, using the above-mentioned cell preservation solution that delays cell growth in cell culture.
[0042] Optionally, the cells are umbilical cord mesenchymal stem cells.
[0043] Optionally, the specific steps of cell preservation include:
[0044] Discard the culture medium of the cells in culture, and add the cell preservation solution to the cells; wherein, the ratio of each milliliter of cell preser...
Embodiment 1
[0069] 1.1. The cell preservation solution is: 1% sodium thiosulfate, 0.1% mannitol, 0.1% sucrose, 10% human albumin and 88.8% phosphate buffer.
[0070] 1.2. The method for cell preservation of the cells in culture: take the umbilical cord mesenchymal stem cells with a confluence of about 60% in culture for cell preservation, discard the original medium, and use 10mL of cell preservation solution per T75 bottle (or 18mL cell preservation solution per T150 bottle, or 20mL cell preservation solution per T175 bottle) take the cell preservation solution; then add the obtained cell preservation solution into the culture bottle, and completely submerge the umbilical cord mesenchymal stem cells; seal the culture bottle with parafilm Air-permeable bottle caps are used to seal and store the cells; then the sealed culture bottles are cooled by a cooling program and then moved to a refrigerator at 4°C for storage. The cooling program includes: first lowering the temperature to 20°C, the...
Embodiment 2
[0073] 2.1. The cell preservation solution is: 2.5% sodium thiosulfate, 0.25% mannitol, 0.05% sucrose, 15% human serum albumin and 82.2% phosphate buffer.
[0074] 2.2. The method for cell preservation of the cells in culture: take the umbilical cord mesenchymal stem cells in culture with a confluence of about 60% for cell preservation, discard the original medium, and use 10mL of cell preservation solution per T75 bottle (or 18mL cell preservation solution per T150 bottle, or 20mL cell preservation solution per T175 bottle) take the cell preservation solution; then add the obtained cell preservation solution into the culture bottle, and completely submerge the umbilical cord mesenchymal stem cells; seal the culture bottle with parafilm Air-permeable bottle caps are used to seal and store the cells; then the sealed culture bottles are cooled by a cooling program and then moved to a refrigerator at 4°C for storage. The cooling program includes: first lowering the temperature to ...
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