The aim of this study was to investigate the biological characteristics of human bone marrow mesenchymal stem cells from bone marrow of different age donors.
The experiments were divided into four groups by donors age:
- group A represented MSC derived from fetal bone marrow;
- group B represented MSC derived from bone marrow of 0-20 years old donors;
- group C represented MSC derived from bone marrow of 20-40 years old donors;
- group D represented MSC derived from bone marrow of donors older than 40.
The growth, purification, proliferation and multipotential abilities of MSC in 4 groups were observed and their immunophenotypes were determined by flow-cytometry.
The level of cytokines (IL-6, SCF, FLT-3L, SDF-1 and TGF-beta1) were assayed by ELISA method.
Cell cycles were analyzed to show the proliferation index (PrI).
MSCs derived from bone marrow of 4 groups were injected subcutaneous into NOD/SCID mouse to observe the safety.
The results showed that different age donors bone marrow all gave rise to MSC.
These cells were similar in morphology, antigenic phenotype, differentiation potential and cell cycle.
The primary culture time of group B was shorter than other groups.
The duration of passage 1 (P1) was 5.5 days, and the duration of P10 was 33 days, after P10 culture, (5.19 ± 2.15)×1010 MSCs were obtained from 8×106 MNC of this group.
The primary culture time of groups A, C, D were longer, the duration of P1 were 15, 7 and 13 days for group A, C and D respectively, and the duration of P10 was 50, 60 and 72 days for group A, C and D, respectively.
After P10 culture, (4.98 ± 2.08)×1010, (1.86 ± 0.47)×1010, (0.64 ± 0.22)×1010 MSCs were obtained from 8×106 MNC of group A, C and D respectively.
The morphology of MSC of group A was longer and slender.
The ability of expansion decreased after P15 for A group, P10 for B group and P8 for C and D groups.
The levels of SCF, FLT3-L, IL-6 and SDF-1 in group B were higher than other groups.
Karyotype analysis showed that MSCs from 4 groups were normal, and tumor-like tissues were not developed after cultured MSCs were inoculated in NOD/SCID mice.
It is concluded that there was relationship between age and the biological characteristics of human bone marrow mesenchymal stem cells.
For clinical use, especially in hematopoietic stem cell transplantation (HSCT), 0-20 years old donors were perfect MSCs donors who can provide sufficient MSCs in relatively short times.
MSCs of group B can be used as stem cell source because the biological characteristics of MSCs of groups B are superior to that of other groups.