Mesenchymal stem cells from cryopreserved human umbilical cord blood
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.
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