Serum-free cryopreservation liquid for neural stem cells
A neural stem cell and stem cell technology, applied in the field of stem cell culture, can solve problems such as pollution, and achieve the effects of overcoming serum pollution, promoting the maintenance of stem cell characteristics, and promoting proliferation
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Embodiment 1
[0027] see figure 1 , technical scheme of the present invention is realized in the following manner, and the step of this experimental method is:
[0028] 1) A neural stem cell cryopreservation solution proposed by the present invention contains the following components in volume percentage: 65%-75% of pre-prepared human neural stem cell serum-free medium, 9-12% of human serum albumin, dimethylformamide A mixed solution of 9-12% sulfoxide, 3-5.5% adenosine, 2.5-5% exosome solution derived from human neural stem cells and 2.5-5% short peptide and polypeptide compound solution derived from human neural stem cells.
[0029] 2) In a specific embodiment, the cryopreservation solution contains human neural stem cell serum-free medium, and the volume percentage can be 65%, 67%, 69%, 71%, 73%, 75%; the volume percentage of human serum albumin can be 9%, 9.5%, 10%, 10.5%, 11%, 12%; the volume percentage of dimethyl sulfoxide can be 9%, 9.5%, 10%, 10.5%, 11%, 12%, the volume of adenosi...
Embodiment 2
[0038] The experimental steps for human nerve cells to be frozen in the above-mentioned freezing solution, and then to be revived, subcultured and expanded are as follows:
[0039] 1. Cell recovery steps:
[0040] a) Quickly take out 1 tube of frozen neural stem cells from the liquid nitrogen tank and place it in an ice box equipped with liquid nitrogen for use;
[0041] b) Turn on the water bath to 37°C;
[0042] c) Use a 10mL pipette to draw 5mL of serum-free medium for neural stem cells into a 15mL centrifuge tube for use;
[0043] d) Take the cells out of the ice box with tweezers, thaw them quickly in a 37°C water bath, and shake gently until there is only a small piece of ice, and the recovery time is controlled within 1 minute;
[0044] e) Use 100-1000μL Tip to suck the recovered cells into the centrifuge tube, shake gently to mix well, the tip can absorb a small amount of cell suspension in the centrifuge tube to wash the cryopreservation tube once, centrifuge at 400...
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