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Isolation and characterization of putative mesenchymal stem cells from mammalian gut

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Abstract

Here, we provide a protocol for reliable isolation and subculture of putative mesenchymal stem cells from mice colons. This method provides a good approach to cultivate and characterize putative colonic mesenchymal stem cells (cMSCs). A high purity of cMSCs can be obtained according to this protocol. The whole isolation processes of cMSCs take about 2 h with two important digestion steps involved. Only with common culture medium, maturation of cMSCs in culture proceeds approximately 2 weeks to allow relevant researches to be conducted. This protocol sheds light on better cultivation of MSCs in vitro from post-natal colon tissues. These putative cMSCs share common phenotypic property with those in vivo reported, and contain potential lineage differentiation capacity. The successful culture of cMSCs in vitro provides an ideal model for study of MSCs biology in the intestine.

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Acknowledgments

This work was supported by the grants from the Natural Science Foundation of Shanghai (No. 12401900704). We thank Stappenbeck and Nick for their sincere assistance and valuable suggestions.

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    Authors and Affiliations

    1. Department of Pharmacy, Changzheng Hospital, The Second Affiliated Hospital of Second Military Medical University, Shanghai, China

      Yulong Tao, Sang Zhu, Hong Yang, Fei Huang & Xia Tao

    2. Department of Pharmacology, School of Pharmacy, Second Military Medical University, Shanghai, China

      Hui Fu

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    1. Yulong Tao
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    2. Sang Zhu
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    3. Hong Yang
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    4. Fei Huang
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    Correspondence to Xia Tao.

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    The authors declare no conflict of interests.

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    Yulong Tao, Sang Zhu and Hong Yang have contributed equally to this work.

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    Tao, Y., Zhu, S., Yang, H. et al. Isolation and characterization of putative mesenchymal stem cells from mammalian gut. Cytotechnology 68, 2753–2759 (2016). https://doi.org/10.1007/s10616-016-9992-z

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