Effect of Cryopreservation Density on Cryopreservation Effect of Peripheral Blood Mononuclear Cells

doi:10.13560/j.cnki.biotech.bull.1985.2018-0646

Abstract

Abstract: The objective of this study is to investigate the effect of cryopreservation density on the cryopreservation of peripheral blood mononuclear cell（PBMC）. Fresh PBMC group（Group F）and 3 PBMC cryopreserved cell density groups：2×10⁷cells/mL（group A）,4×10⁷ cells/mL（group B）,6×10⁷cells/mL（group C）were set up. By comparing these data,such as the survival rate of PBMC before and after cryopreservation,as well as the amplification multiples,lymphocyte subsets,and in vitro killing efficiency of multiple cytokines-induced killer（CIK）cells after resuscitation,the cryopreservation effect was validated. Results shows as,the cell viability of group F,group A,group B and group C before and after cryopreservation were（96 + 0.3）%,（95.6 + 0.4）%,（94.7 + 0.2）% and（94.9 + 0.4）%,respectively,i.e.,the cell viability of group B and group C were significantly lower than that of group A（P<0.05）. At 14 days after cell amplification,the cell amplification of group F and group A,group B,and group C were（156.4 + 18.2）times,（160.2 + 28.4）times,（126.1 + 19.8）times,and（110.4 + 11.3）times,respectively,i.e.,that cell amplification of group B and group C was significantly lower than that of group F（P < 0.05）. There was no significant difference in the results of lymphocyte subsets before PBMC freezing and after PBMC resuscitation. Compared with group F,at 14 d after cell amplification of group A,group B,and group C,the lymphocyte subsets and the in vitro killing efficiency of CD3+,CD3+CD4+,CD3+ CD8+,CD3-CD56+ and CD3+CD56+ demonstrated no significant difference（P>0.05）. In conclusion,too high cryopreservation density may affect the cryopreservation effect of PBMC so as to indirectly affect the cell resuscitation. However,the too low cryopreservation density increases the cryopreservation volume so as to greatly raise cost. Therefore,choosing appropriate cell cryopreservation density is an issue that must be considered for a cell library or cell bank.

Key words: cryopreservation, peripheral blood mononuclear cell, survival rate, cell density

LIN Ke-jia, LIU Fu-ping, MA Dong-lei, HU Rui, WEI Zong-ke, TAN Yi. Effect of Cryopreservation Density on Cryopreservation Effect of Peripheral Blood Mononuclear Cells[J]. Biotechnology Bulletin, 2019, 35(6): 119-124.

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