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The effect of extended passaging on the phenotype and osteogenic potential of human umbilical cord mesenchymal stem cells

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Abstract

Retaining biological characteristics in the extended passaging is crucial for human umbilical cord mesenchymal stem cells (hUCMSCs) in tissue engineering. We aimed to assess morphology, viability, MSC marker expression, and osteogenic activity of hUCSMCs after extended passaging. Passages 4 (P4) and 16 (P16) hUCMSCs displayed similar morphology and viability. The flow cytometry results showed that CD73, CD90, and CD105 were highly expressed at P1–P16. CD166 expression decreased progressively from 90 % at P2 to 61.5 % at P5 (p < 0.05), followed by stable expression through P16. Results from calcium deposition alkaline phosphatase activity and RT-PCR assay showed that both P4 and P16 hUCMSCs differentiated down an osteogenic lineage, with no significant difference in osteogenic capacity (p < 0.05). High-passage UMCSCs maintained stable expression of MSC CD markers as well as stable osteogenic activity. hUCMSCs may thus be suitable for tissue engineering and regenerative medicine applications.

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Acknowledgments

We gratefully acknowledge Prof. Zhengliang Chen at the Southern Medical University for FCM assistance and useful discussions. This study was supported by National Natural Science Foundation of China 31328008 (LZ), 31100695(LZ) Natural Science Foundation of Guangdong s20130010014253 (LZ), Guangdong Provincial Science and Technology Project 2012B010200024 (LZ) and Guangzhou Science and Technology Project 2012027(LZ).

Conflict of Interest

None declared.

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    Authors and Affiliations

    1. Department of Orthopaedic Surgery, The Third Affilliated Hospital, Southern Medical University, Guangzhou, China

      Zhe Shi

    2. Department of Orthopaedic Surgery, Nanfang Hospital, Southern Medical University, 1838 North Guangzhou Ave, Guangzhou, 510515, China

      Liang Zhao, Gengtao Qiu & Ruixuan He

    3. Department of Orthopaedic Surgery, ShunDe First People Hospital, Shunde, Foshan, China

      Gengtao Qiu

    4. Department of Chemical & Petroleum Engineering, University of Kansas, 4132 Learned Hall, 1530 W 15th Street, Lawrence, KS, 66045, USA

      Michael S. Detamore

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    1. Zhe Shi
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    Correspondence to Liang Zhao or Michael S. Detamore.

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    Liang Zhao, Zhe Shi and Gengtao Qiu have contributed equally to this work.

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    Shi, Z., Zhao, L., Qiu, G. et al. The effect of extended passaging on the phenotype and osteogenic potential of human umbilical cord mesenchymal stem cells. Mol Cell Biochem 401, 155–164 (2015). https://doi.org/10.1007/s11010-014-2303-0

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