Method for freezing and thawing placental whole cells and separating and expanding stem cells
A whole-cell, qualitative stem cell technology, applied in the field of placental whole cell cryopreservation, resuscitation, isolation and expansion of stem cells, can solve the problem that the method and technology are not fully mature
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Embodiment 1
[0113] Embodiment 1, placental whole cell cryopreservation, resuscitation and the method of stem cell separation and expansion after resuscitation Law
[0114] The placental tissue cryopreservation method includes the following steps:
[0115](1) Placental tissue cleaning: The placental tissue is processed in a biological safety cabinet. According to the size of the placenta, an appropriate amount of PBS buffer is used to wash the placental tissue 2-3 times, so that the residual blood on the surface of the placental tissue is washed clean, and there is no blood clot on the surface of the placenta. piece;
[0116] (2) Digestion of placenta tissue: use surgical scissors to cut placental leaflets from the placental tissue obtained in step (1), transfer the leaflets to a petri dish, add 25ml PBS buffer solution and cut the placental leaflets as much as possible, add 25ml 0.25 % trypsin (Gibco) (the volume ratio of trypsin to PBS buffer is 1:1) and mix the tissue, put the cultu...
Embodiment 2
[0133] Embodiment 2, placental whole cell cryopreservation, resuscitation and the method of stem cell separation and expansion after resuscitation Law
[0134] Carried out with reference to the method of Example 1. The recovered placental whole cells began to have adherent cells on the 7th day of culture, and the cell fusion rate reached 80% by the 19th day of culture. After 3 passages, the cell purity was greater than 90%.
Embodiment 3
[0135] Embodiment 3, placental whole cell cryopreservation, resuscitation and the method of stem cell separation and expansion after resuscitation Law
[0136] Carried out with reference to the method of Example 1. The recovered placental whole cells began to have adherent cells on the 8th day of culture, and the cell fusion rate reached 80% by the 21st day of culture. After 3 passages, the cell purity was greater than 85%.
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