Effect of age and passage on canine bone marrow derived mesenchymal stem cells

摘要

Mehrabani D, Khodakaram-Tafti A, Asadi-Yousefabad SL, Dianatpour M, Zare S, Tamadon A, Nikeghbalian S, Jahromi AR, Ahmadlou S., Effect of age and passage on canine bone marrow derived mesenchymal stem cells, Onl J Vet Res., 19(10): 663-671, 2015. Bonemarrow mesenchymal stem cells (BM-MSCs) from proximal humerus of mixed breed healthy dogs were isolated for culture. BM-MSCs of 4, 5, and 8th passage from a 3yr old dog and 8th passage from a 1yr old dog were seeded in 12 plates at a density of 5x104 cells per well for 8 days. Three wells per day were used to determine cell numbers. Morphology, growth curves and population doubling time (PDT) for each passage were calculated and cell viability after thawing was evaluated. BM-MSCs displayed fibroblastic-like morphology during all passages. RT-PCR showed that cells were positive for MSC marker CD90 but negative for haematopietic markers CD34 and CD45. PDT during the 8th passage in the 1yr dog was 41.1h whereas during the 4, 5 and 8th passage in the 3yrdog, were 65.8, 55.3 and 90h, respectively. Growth and proliferation of BM-MSCs in the 8th passage in 1yr dog was greater compared with older dog (P<0.01) except for day 3 when there was no difference (P>0.01). Cell viability was 81.6% for the 3rd passage in the 1yr dog and 86.6% during the 1st passage in the 3yr dog. BM-MSCs were spindle-shaped in all passages. Results suggest that the increase in PDT occurs with increased age as no differences were shown between the mean numbers of BM-MSCs during different passages.