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OJVRTM

Online Journal of Veterinary Research©

Volume 19(10): 663-671, 2015.


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

 

Davood Mehrabani1, Azizollah Khodakaram-Tafti2, Seyedeh Leili Asadi-Yousefabad3,*, Mehdi Dianatpour1,4, Shahrokh Zare1,

Amin Tamadon1, Saman Nikeghbalian5, Alireza Raayat Jahromi6, Somayeh Ahmadlou1

 

1Stem Cell and Transgenic Technology Research Center, Shiraz University of Medical Sciences, 2Department of Pathobiology Sciences, School of Veterinary Medicine, Shiraz University, 3Young Researchers and Elite Club, Yasuj Branch, Islamic Azad University, Yasuj, 4Department of Medical Genetics, School of Medicine, Shiraz University of Medical Sciences, 5Shiraz Organ Transplantation Research Center, Namazi Hospital, Shiraz University of Medical Sciences, 6Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

 

ABSTRACT

 

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. Bone marrow 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  5×104 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 hematopoietic 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 3yr dog, 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. Results should be viewed with caution considering the fact that only 2 dogs were tested.

 

KEY-WORDS: Mesenchymal stem cell, Bone marrow, Isolation, Culture, Characterization, Dog.


 

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