Cell cryopreservation fluid and application thereof
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A technology for freezing liquid and cells, applied in the field of biomedicine, can solve the problem of low cell viability, achieve high cell viability, avoid virus contamination, and be widely applicable
Inactive Publication Date: 2019-01-18
GUANGDONG CHINAHEALTH LIFE SCI CO LTD
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[0004] The invention provides a cell cryopreservation solution and its application, which solves the technical problem that the cells frozen in the existing cell cryopreservation system are not high in viability after resuscitation
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Embodiment 1
[0043] 1) Prepare cell cryopreservation solution:
[0044] ① Configure by volume fraction (V%): DMSO 30%, human albumin 10%-20%, low molecular dextran 10%-20%, human mesenchymal stem cell exosomes 20%, hydroxyethyl starch 10 %-20%, the balance is RPMI 1640 basal medium.
[0045] 2) Obtain endometrial stem cells
[0046] References: Zhou Hua, Song Lina, Zhang Min, et al. Isolation, culture and identification of human endometrial stem cells in vitro, Chinese Journal of Traditional Chinese Medicine [J], 2014, 32(8): 1907~1908. Uterus is obtained by the method Primary cells of endometrial stem cells are subcultured to obtain cells from P2 to P5.
[0047] 3) Specific freezing storage steps:
[0048] After resuspending in PBS, centrifuge at 1500r / min for 5min, and discard the supernatant. Take 50uL of cell suspension, mix it according to cell suspension (v): 0.4% trypan blue (v) = 1:1, then calculate cell viability and number; use the cell jelly prepared in step 1) of this example. Adjust...
Embodiment 2
[0054] 1) Prepare cell cryopreservation solution: 10% DMSO, 5%-10% human albumin, 5%-10% low molecular dextran, 20% exosomes, 20% hydroxyethyl starch, the balance is RPMI 1640 basal medium.
[0055] The operation steps from step 2) to step 6) are the same as in Example 1.
Embodiment 3
[0057] 1) Prepare cell cryopreservation solution: 10% DMSO, 10%-20% human albumin, 10%-20% low molecular dextran, 20% exosomes, 20% hydroxyethyl starch, the balance is RPMI 1640 basal medium.
[0058] The operation steps from step 2) to step 6) are the same as in Example 1.
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Abstract
The invention relates to the technical field of biomedicine, in particular to cell cryopreservation fluid and application thereof. The cell cryopreservation fluid disclosed by the invention is prepared from the following components: dimethyl sulfoxide, human albumin, low-molecular dextran, a human mesenchymal stem cell exosome, hydroxyethyl starch and a basal culture medium. Cells cryopreserved and revived by using the cell cryopreservation fluid are high in viability, the morphologic change is not large, differentiation cannot be caused during culture, and surface antigen detection accords with the standard; in addition, by optimizing a cell cryopreservation scheme, death of a large amount of cells caused by ice crystal formation during a cryopreservation process of the cells can be avoided as much as possible, a cryopreservation system is optimized, revived cells can be directly fed back to a human body, and animal derived serum is not added, animal derived virus infection is avoided; the cell cryopreservation fluid is wide in application, and the technical problem that the cell viability after the reviving of the cells which are cryopreserved by an existing cell cryopreservationsystem is not high is solved.
Description
Technical field [0001] The invention relates to the field of biomedicine technology, in particular to a cell cryopreservation liquid and its application. Background technique [0002] Cryopreservation of cells is one of the main methods of cell preservation. Using cryopreservation technology to store the cells in liquid nitrogen at -196°C at low temperature can temporarily remove the cells from the growth state and preserve their cell characteristics, avoiding senescence and differentiation of the cells during the culture process. In addition, proper preservation of a certain amount of cells can prevent the cells in culture from being contaminated or other accidents that would cause the cells to be lost, which plays a role in cell preservation. In addition, you can also use the form of cell cryopreservation to purchase, gift, exchange and transport certain cells. [0003] When the cells are cryopreserved, add a protective agent-glycerol or dimethyl sulfoxide (DMSO) with a final c...
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