Research Article

Isolation, Culture, Identification and Differentiation of Canine Amniotic Mesenchymal Stem Cells in Vitro  

Yukun Zhang1 , Liheng Bai2 , Yan Li1 , Moning Liu1 , Guifang Cao1 , Dafu Zhi1
1 Animal Tissue Embryology and Developmental Biology Laboratory, College of Life Sciences, Inner Mongolia Agricultural University, Huhhot, China
2 Inner Mongolia Maternal and Child Health Hospital, Huhhot, China
Author    Correspondence author
Animal Molecular Breeding, 2017, Vol. 7, No. 1   doi: 10.5376/amb.2017.07.0001
Received: 20 Apr., 2017    Accepted: 25 Apr., 2017    Published: 28 Apr., 2017
© 2017 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Zhang Y.K., Bai L.H., Li Y., Liu M.N., Gao G.F., and Zhi D.F., 2017, Isolation, culture, identification and differentiation of canine amniotic mesenchymal stem cells in vitro, Animal Molecular Breeding, Vol.7, No.1, 1-13 (doi: 10.5376/amb.2017.07.0001)

Abstract

Recent years, people are paying more and more attention to amniotic membrane tissue which has a rich resource and no ethical restrictions. Canine amniotic mesenchymal stem cells (cAMSCs) were collected by enzyme digestion method from canine amniotic membrane tissues. The cell morphology was continuously observed at different passages and cell growth curves of passage 3 and passage 9 were drawed. Stem cell related proteins and genes were identified by immunofluorescence and RT-PCR. The multiple differentiation potential of cAMSCs was identified by osteogenesis and adipogenesis. This study showed that the cAMSCs was long fusiform and could be continuously subcultured in vitro. The immunofluorescence results showed that Vimentin and SSEA-4 were positively expressed. The RT-PCR results showed that stem cell related genes OCT4, SOX2 and NANOG were positively expressed too. The cAMSCs could be induced to osteoblasts and adipocytes in vitro. The markers associated with osteogenesis and adipogenesis, such as COL1A1 and LPL, were positively expressed after differentiation. The results showed that the cAMSCs could be successfully isolated and cultured in vitro. Stem cell related proteins and genes were positively expressed. And the cAMSCs has the multiple differentiation potential. This study showed that cAMSCs could be a rich source of stem cells in veterinary. Furthermore, cAMSCs may be useful as a cell therapy application for veterinary regenerative medicine.

Keywords
Canine; Amnion; Mesenchymal stem cells; Differentiation; Veterinary cell therapy
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