Objective: To investigate the expression of human leukocyte antigens (HLA) in human fetal bone marrow mesenchymal stem cells (MSCs) after long time culture as well as the changes in response to interferon-gamma (IFN-gamma) treatment.
Methods: Human fetal MSCs were collected from 23 to 24-weeks-old fetues with the approval of Ethic Committee of Peking University Health Science Center. The cells of passage 5 and passage 12 were analyzed for HLA expression before and after IFN-gamma (50 microg/L) treatment by flow cytometry at different time points. The passage 5 cells were also treated with an additional dose of IFN-gamma (5 microg/L) and the HLA expression was analyzed 24, 48, 72, 96 and 120 h after treatment. RT-PCR was used to evaluate HLA-G and HLA-E expression at mRNA level in human fetal MSCs.
Results: Flow cytometry results showed that fetal MSCs expressed high level of HLA class I (HLA-I) antigen, but extremely low level of HLA class II (HLA-II) antigen. The percentage of HLA-I positive MSCs in total MSCs was approximately over 50%, while that of HLA-II positive MSCs in total MSCs was less than 10%. IFN-gamma (50 microg/L) enhanced the HLA-I and HLA-II expression in a time dependent manner and increased the percentage of HLA-I positive cells in both passage 5 and passage 12 cells but preferably in passage 5 cells. The enhanced HLA-II expression was seen 48 h after IFN-gamma treatment in passage 5 cells (59.9%) but 72 h in passage 12 cells (48.1%). The treatment of 5 microg/L IFN-gamma also increased percentage of HLA-I and HLA-II positive MSCs, but with a relatively less extent compared to the 50microg/L IFN-gamma treatment group. RT-PCR result indicated HLA-G and HLA-E were expressed at mRNA level in human fetal MSCs.
Conclusion: Human fetal MSCs can be induced to express both HLA class I and class II antigens by IFN-gamma. The long time culture might reduce the IFN-gamma effects. Human fetal MSCs express HLA-G and HLA-E at mRNA level.