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Korean Journal of Obstetrics & Gynecology 2006;49(5):1073-1084.
Published online May 1, 2006.
Isolation and characterization of mesenchymal stem cells derived from umbilical cord blood.
Myung Hee Kim, Jin Hyun Park, Chan Soo Shin, Hee Joong Kim, Jung Gu Kim
1Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Korea. kimjg@.snu.ac.kr
2Department of Internal Medicine, Seoul National University, Korea.
3Department of Orthopedic Surgeon, Seoul National University, Korea.
Abstract
OBJECTIVE
The purposes of this study were to isolate and characterize mesenchymal stem cells (MSCs) with osteogenic and adipogenic potential from umbilical cord blood (UCB). METHODS: MSCs were isolated using a density gradient centrifugation and extensive subcultivation from UCB. The proliferation capability, cell cycle, cytochemical markers, and immunophenotype of these MSCs were measured. The transcript of osteoblast-specific markers, alkaline phosphatase (ALP), and calcium deposit were analyzed from MSCs cultured in osteogenic media for 1-4 weeks, and MSCs exposed in adipogenic media were stained by Oil red after 4 weeks of culture. RESULTS: More than 84% of MSCs were in the G0/G1 phase of cell cycle. MSCs were positive for periodic acid-Schiff, and alpha-naphthyl acetate esterase activity, but negative for sudan black-B, and ALP activities. MSCs expressed CD13, CD29, CD 44, CD49e, CD51, CD54, CD 90, SH2, and SH3, but did not express CD11b, CD14, CD31, CD34, CD45, CD49d, CD106, CD133, CD144, CD146, CD163, CD166, and Stro-1. When MSCs were cultured in osteogenic media for 1-4 weeks, they expressed transcripts of osteoblastic specific markers: Runx-2, ALP, and procollagen type I. During culture of 2-4 weeks, ALP activity was detected and quantification increased significantly, and the deposition of a calcified matrix became evident. Exposure of MSCs to adipogenic media resulted in morphological change stained by Oil Red O. CONCLUSION: These data indicate that UCB contains MSCs with osteogenic and adipogenic differentiation potential, and may serve as an potential source of MSCs to be utilized in cell therapy for various diseases.
Key Words: Umbilical cord blood, Mesenchymal stem cell, Characteristics


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