Objective: To investigate the effect of allogeneic bone on viability and proliferation of human peripheral blood lymphocyte by CCK-8 method and CFSE method. Methods: The lymphocyte viability was detected by CCK-8 method after co-culture of human fresh peripheral blood lymphocytes and the sample extraction (decalcified bone matrix and freeze-dried tissue engineered bone). Also, the lymphocyte proliferation was determined by flow cytometry after co-culture of fresh peripheral blood lymphocytes labeled with CFSE and the sample extraction. Results: The result of CCK-8 method showed that there was no significant difference in absorbance value between decalcified bone matrix and medium control. But there was a significant increase of absorbance value in freeze-dried tissue engineered bone compared with that in medium control (P<0.05), suggesting that freeze- dried tissue engineered bone improved lymphocyte viability. The result of CFSE method showed that there was no significant difference in Dil, PF, EI and RI of lymphocyte, and Dil of lymphocyte subtype between freeze-dried tissue engineering bone and medium control (P>0.05). The results suggested that freeze-dried tissue engineered bone had no significant effect on the proliferation of human peripheral blood lymphocyte. Conclusion: Compared with medium control, decalcified bone matrix has no significant effect on human peripheral blood lymphocyte viability. Freeze-dried tissue engineered bone improves the human peripheral blood lymphocyte viability, but dose not significantly affect the proliferation of human peripheral blood lymphocyte. CCK-8 method mainly reflects the cell viability, but CFSE method could reflect the cell proliferation more reasonable.
SHAO An-liang, MU Yu-feng, CHEN Li-yuan, CHEN Liang, XU Li-ming
. Evaluation of allogeneic bone on viability and proliferation of human peripheral blood lymphocyte in vitro*[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(9)
: 1505
-1510
.
DOI: 10.16155/j.0254-1793.2022.09.03
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