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Isolation and characterization of mesenchymal stem cells from human umbilical cord blood: Reevaluation of critical factors for successful isolation and high ability to proliferate and differentiate to chondrocytes as compared to mesenchymal stem cells from bone marrow and adipose tissue

Authors

  • Xiaohong Zhang,

    1. Cell Therapy Research and Development Laboratory, New York Blood Center, New York, New York
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    • Xiaohong Zhang, Masako Hirai, Tsuneo A. Takahashi's former address is Division of Cell Processing, Facility of Cell Processing and Cryopreservation, Tokyo Cord Blood Bank, The Institute of Medical Science, The University of Tokyo 4-6-1, Shirokanedai, Minato-ku Tokyo 108-8639, Japan.

  • Masako Hirai,

    1. Cell Therapy Research and Development Laboratory, New York Blood Center, New York, New York
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    • Xiaohong Zhang, Masako Hirai, Tsuneo A. Takahashi's former address is Division of Cell Processing, Facility of Cell Processing and Cryopreservation, Tokyo Cord Blood Bank, The Institute of Medical Science, The University of Tokyo 4-6-1, Shirokanedai, Minato-ku Tokyo 108-8639, Japan.

  • Susana Cantero,

    1. Cell Therapy Research and Development Laboratory, New York Blood Center, New York, New York
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  • Rodica Ciubotariu,

    1. National Cord Blood Program, New York Blood Center, New York, New York
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  • Ludy Dobrila,

    1. National Cord Blood Program, New York Blood Center, New York, New York
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  • Allen Hirsh,

    1. Cryobiophysica, Inc., Silver Spring, Maryland
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  • Koichi Igura,

    1. Department of Regulatory Science, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan
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  • Hitoshi Satoh,

    1. Laboratory of Tumor Cell Biology, Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan
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  • Izuru Yokomi,

    1. Department of Pharmacology, School of Medicine, St. Marianna University, Kawasaki, Japan
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  • Toshihide Nishimura,

    1. Department of Immunology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan
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  • Satoru Yamaguchi,

    1. Yamaguchi Hospital, Funabashi, Chiba, Japan
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  • Kotaro Yoshimura,

    1. Department of Plastic Surgery, The University of Tokyo, School of Medicine, Tokyo, Japan
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  • Pablo Rubinstein,

    1. National Cord Blood Program, New York Blood Center, New York, New York
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  • Tsuneo A. Takahashi

    Corresponding author
    1. Cell Therapy Research and Development Laboratory, New York Blood Center, New York, New York
    • Cell Therapy Research and Development Laboratory, New York Blood Center, 310E. 67th Street, New York, NY 10065.
    Search for more papers by this author
    • Xiaohong Zhang, Masako Hirai, Tsuneo A. Takahashi's former address is Division of Cell Processing, Facility of Cell Processing and Cryopreservation, Tokyo Cord Blood Bank, The Institute of Medical Science, The University of Tokyo 4-6-1, Shirokanedai, Minato-ku Tokyo 108-8639, Japan.


  • Author Contributions: X.Z. and T.T. designed research; X.Z., M.H., S.C., K.I., and T.T. performed research; R.C. and S.Y. were responsible for cord blood collection; A.H. performed statistical analysis; H.S. and I.Y. performed karyotype analysis; T.N. performed the immunosuppressive study; K.Y. was responsible for adipose tissue collection; L.D. provided advice on cord blood processing; P.R. advised on the manuscript: and X.Z. and T.T. wrote the paper.

  • Conflict-of-interest disclosure: The authors declare no competing financial interests.

Abstract

Human umbilical cord blood (CB) is a potential source for mesenchymal stem cells (MSC) capable of forming specific tissues, for example, bone, cartilage, or muscle. However, difficulty isolating MSC from CB (CB-MSC) has impeded their clinical application. Using more than 450 CB units donated to two public CB banks, we found that successful cell recovery fits a hyper-exponential function of time since birth with very high fidelity. Additionally, significant improvement in the isolation of CB-MSC was achieved by selecting cord blood units having a volume ≥90 ml and time ≤2 h after donor's birth. This resulted in 90% success in isolation of CB-MSC by density gradient purification and without a requirement for immunoaffinity methods as previously reported. Using MSC isolated from bone marrow (BM-MSC) and adipose tissue (AT-MSC) as reference controls, we observed that CB-MSC exhibited a higher proliferation rate and expanded to the order of the 1 × 109 cells required for cell therapies. CB-MSC showed karyotype stability after prolonged expansion. Functionally, CB-MSC could be more readily induced to differentiate into chondrocytes than could BM-MSC and AT-MSC. CB-MSC showed immunosuppressive activity equal to that of BM-MSC and AT-MSC. Collectively, our data indicate that viable CB-MSC could be obtained consistently and that CB should be reconsidered as a practical source of MSC for cell therapy and regenerative medicine using the well established CB banking system. J. Cell. Biochem. 112: 1206–1218, 2011. © 2011 Wiley-Liss, Inc.

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