Isolation of mesenchymal stromal/stem cells from cryopreserved umbilical cord blood cells

  • Fujii Sumie
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital Department of Hematology/Oncology, Graduate School of Medicine, Kyoto University
  • Miura Yasuo
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital
  • Iwasa Masaki
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital Division of Gastroenterology and Hematology, Shiga University of Medical Science
  • Yoshioka Satoshi
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital
  • Fujishiro Aya
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital Division of Gastroenterology and Hematology, Shiga University of Medical Science
  • Sugino Noriko
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital Department of Hematology/Oncology, Graduate School of Medicine, Kyoto University
  • Kaneko Hitomi
    Department of Hematology, Osaka Red Cross Hospital
  • Nakagawa Yoko
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital
  • Hirai Hideyo
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital
  • Takaori-Kondo Akifumi
    Department of Hematology/Oncology, Graduate School of Medicine, Kyoto University
  • Ichinohe Tatsuo
    Department of Hematology and Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima University
  • Maekawa Taira
    Department of Transfusion Medicine and Cell Therapy, Kyoto University Hospital

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Abstract

<p>Umbilical cord blood (UCB) has advantages over other tissues because it can be obtained without an invasive procedure and complex processing. We explored the availability of cryopreserved UCB cells as a source of mesenchymal stromal/stem cells (MSCs). MSCs were successfully isolated from six of 30 UCB units (median volume, 34.0 mL; median nucleated cell number, 4.4×108) that were processed and cryopreserved using CP-1/human serum albumin. This isolation rate was lower than that (57%) from non-cryopreserved UCB cells. The number of nucleated cells before and after hydroxyethyl starch separation, UCB unit volume, and cell viability after thawing did not significantly differ between UCB units from which MSCs were successfully isolated and those from which they were not. When CryoSure-DEX40 was used as a cryoprotectant, MSCs were isolated from two of ten UCB units. Logistic regression analysis demonstrated that the cryopreservation method was not significantly associated with the success of MSC isolation. The isolated MSCs had a similar morphology and surface marker expression profile as bone marrow-derived MSCs and were able to differentiate into osteogenic, adipogenic, and chondrogenic cells. In summary, MSCs can be isolated from cryopreserved UCB cells. However, the cryopreservation process reduces the isolation rate; therefore, freshly donated UCB cells are preferable for the isolation of MSCs.</p>

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