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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
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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