Mesenchymal stem cells from cryopreserved human umbilical cord blood

https://doi.org/10.1016/j.bbrc.2004.04.206Get rights and content

Abstract

Umbilical cord blood (UCB) is well known to be a rich source of hematopoietic stem cells with practical and ethical advantages, but the presence of mesenchymal stem cells (MSCs) in UCB has been disputed and it remains to be validated. In this study, we examined the ability of cryopreserved UCB harvests to produce cells with characteristics of MSCs. We were able to obtain homogeneous plastic adherent cells from the mononuclear cell fractions of cryopreserved UCB using our culture conditions. These adherent cell populations exhibited fibroblast-like morphology and typical mesenchymal-like immunophenotypes (CD73+, CD105+, and CD166+, etc.). These cells presented the self-renewal capacity and the mesenchymal cell-lineage potential to form bone, fat, and cartilage. Moreover, they expressed mRNAs of multi-lineage genes including SDF-1, NeuroD, and VEGF-R1, suggesting that the obtained cells had the multi-differentiation capacity as bone marrow-derived MSCs. These results indicate that cryopreserved human UCB fractions can be used as an alternative source of MSCs for experimental and therapeutic applications.

Section snippets

Materials and methods

Samples: collection and cryopreservation. The Institutional Review Board of Ajou University Hospital approved this study and all samples were obtained with informed consent. Forty BM and 47 UCB samples which had undergone cryostorage for 0.1–5 years were used for the study.

Before freezing, UCB cells were separated by the Ficoll–Hypaque (Histopaque-1077; Sigma, MO, USA) density-gradient method and washed with Dulbecco’s phosphate-buffered saline (DPBS; HyClone, UT, USA). Separated MNCs were

Isolation and morphological analysis of cryopreserved UCB-derived mesenchymal stem cells

The duration of storage in frozen state for UCB was 0.1–5 years, and there were no differences in viability depending on the storage duration. When thawed, both BM- and UCB-derived cells were recovered with more than 90% viability. Frozen UCB-derived mononuclear cells were plated at a density of 3 × 105 cells/cm2 and formed adherent heterogeneous cell populations after 4–7 days in culture, which consisted of round and spindle-shaped cells. In the initial passage of culture, the cells proliferated

Discussion

UCB as the power of HSCs has been increasingly used for various clinical settings since 1988 [17]. Since then, hundreds of thousands of UCB collections have been frozen and stored throughout the world, in anticipation of their potential use to treat various disorders. Thus, study on the long-term storage of UCB-derived stem cells including HSCs and MSCs is of critical importance. We have investigated whether MNC fractions from cryopreserved UCB contained stem cells which represented typical

Acknowledgements

This research was supported by a Grant (SC-13100) from Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology, Republic of Korea.

References (22)

  • S. Wakitani et al.

    Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine

    Muscle Nerve

    (1995)
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