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Isolation, culture, and induced multiple differentiation of Mongolian sheep bone marrow-derived mesenchymal stem cells

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An Erratum to this article was published on 27 March 2015

Abstract

The aim of this paper was to explore the optimal method of isolating, purifying, and proliferating Mongolian sheep bone marrow-derived mesenchymal stem cells (BMSCs) and their multiple differentiation potentialities. Bone marrow (BM) was punctured from ∼1-year-old sheep, and BMSCs were harvested through gradient centrifuge and adherent cultures. Analysis of the growth of the passage 1, 5, and 10 cultures revealed an S-shaped growth curve with a population doubling time of 31.2 h. Karyotyping indicated that the chromosome number in the Mongolian sheep was 2n = 54, comprising 26 pairs of autosomes and one pair of sex chromosomes (XY). RT-PCR demonstrated that OCT4, SOX2, and Nanog genes at passage 3 were positively expressed. The P3 BMSCs were cultured in vitro under inductive environments and induced into adipocytes, osteoblasts, chondrocytes, neural cells, and cardiomyocytes. Their differentiation properties were confirmed by histological staining, such as oil red, Alizarin red, hematoxylin–eosin, toluidine blue, and periodic acid schiff. RT-PCR showed that the specific genes to be induced were all expressed. This proves that the isolated cells are indeed the BMSCs and also provides valuable materials for somatic cell cloning and transgenic research.

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Acknowledgments

The authors would like to thank Professor Huanmin Zhou for providing the Mongolian sheep and Dr. Yiyi Liu and Lu Li for their technical help. This study was supported by a Grant-in-Aid for the major project of the Inner Mongolia Natural Science Foundation (A) (2012ZD03), a Grant-in-Aid for China Agricultural University cooperation project (B) (2010), and Inner Mongolia Key Laboratory of Biomanufacture.

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Authors and Affiliations

  1. College of Life Sciences, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot, Inner Mongolia Autonomous Region, 010018, China

    Zongzheng Liu, Wei Wang, Jinfang Gao, Huanmin Zhou & Yanru Zhang

  2. Key Laboratory of Biological Manufacturing of Inner Mongolia Autonomous Region, Hohhot, China

    Zongzheng Liu, Wei Wang, Jinfang Gao, Huanmin Zhou & Yanru Zhang

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  1. Zongzheng Liu
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  2. Wei Wang
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  4. Huanmin Zhou
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Correspondence to Yanru Zhang.

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Editor: T. Okamoto

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Liu, Z., Wang, W., Gao, J. et al. Isolation, culture, and induced multiple differentiation of Mongolian sheep bone marrow-derived mesenchymal stem cells. In Vitro Cell.Dev.Biol.-Animal 50, 464–474 (2014). https://doi.org/10.1007/s11626-013-9725-y

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