| Objective:To evaluate the age related changes of T lymphocyte subsets in C57BL/6 mice and splenic T cells proliferative ability.Fetus-derived mesenchymal stem cells were transplanted to the aged mice and the improvements of T lymphocyte subsets and splenic T cells proliferative ability were detected to assessment the anti-aging effects.Methods:Multi-color immunofluorescence techniques were used to analysis relative numbers of T lymphocyte subsets include CD4+,CD8+,naive and memory CD4+ and CD8+,CD8+CD28+T cells in peripheral blood of C57BL/6 mice from different age group(Group I: 2 months old;Group II: 7 months old;Group III: 21 months old).Splenocytes isolated from different group mice were stimulated with Con A to evaluate the proliferative ability.Cells isolated from mice fetus and identified by biomarker of cells and differentiation ability.21 months old mice were given stem cell therapy,1×106 cells were transplanted per week for 3 times,a month later,the T lymphocyte subsets and proliferative ability were detected to evaluate the effect.Results:1.The percentage of CD4+ T cells from group I,the group II and group III was4.08%,49.71% and 47.22%,The percentage of CD8+T cells from group I,the group II and group III was 33.18%,46.00% and 51.10%.Compared with group I,the group II and group III had a significant reduction in the the percentage of CD4+ T cells and increase in the percentage of CD8+T cells in peripheral blood(P < 0.05);while compared between group II and group III,there was not a significant different in the percentage of CD4+ and CD 8+ T cells(P>0.05).2.The percentage of naive CD4+T cells from group I,the group II and group III was 21.68%,7.44% and 10.73%,The percentage of memory CD4+ T cells from group I,the group II and group III was 8.73%,17.35% and 37.47%,the percentage of naive CD8+T cells from group I,the group II and group III was 43.83%,27.36% and 13.92%,The percentage of memory CD8+ T cells from group I,the group II and group III was9.10%,11.31% and 32.10%.Compared with group I,the group II and group III had a significant reduction in the the percentage of naive CD4+T cells and naive CD8+T cells(P<0.05);the group III had a significant increase in the percentage of memoryCD4+ T cells and memory CD8+ T cells(P<0.05).Compared with group II,the group III had a significant reduction in the percentage of naive CD8+ T cells(P<0.05)and significant increase in the percentage of memory CD4+T cells and memory CD8+T cells(P<0.05).3.The percentage of CD8+CD28+ T cells from group I,the group II and group III was 24.38%?31.64%?47.67%.Compared with group I and group II,the Igroup III had a significant increase in the percentage of CD8+CD28+ T cells(P<0.05).4.The T lymphocyte proliferation in vitro that group II and group III had a lower proliferative capacity than group I,Between group II and group III(P<0.05),there was not a significant difference(P>0.05).5.Fetus-derived mesenchymal stem cells' expression of CD29 was 99.9%,CD44 was 90.3%,SCA-1 was 85%,CD34 was 2.9%,CD45 was 0.5%,CD11 b was 0.5%.Cells can be induced to differentiate into osteogenic and adipogenic cell types.6.The T lymphocyte subsets of treated group mice: the percentage of CD4+ and CD8+T cells was 53.12% and 46.36%,the percentage of naive and memory CD4+T cells was 5.84% and 53.94%,the percentage of CD8+CD28+ T cells was 42.28%.Compared with Control group(group III),there were not significant difference in the T lymphocyte subsets(P>0.05).7.Compared with Control group(group III),there was not a significant difference in T lymphocyte proliferation(P>0.05).Conclusions:1.There were age-related changs in the percentages of CD4+,CD8+,naive and memory CD4+ and CD8+ T cells from the peripheral blood of C57BL/6 mice and those index can be used to evaluate the immunosenescence of C57BL/6 mice.2.The cells we got were mesenchyma stem cells.3.Stem cell therapy may not improve the T lymphocyte subsets and proliferative ability of 19 months. |
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