Isolation and Characterization of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood Mononuclear Cells

Chih-Yang Chang ； Po-Han Chen ； Chia-Jung Li ； Shang-Chieh Lu ； Yu-Chun Lin ； Po-Huang Lee ； Ying-Hsien Kao

《E-Da Medical Journal》 3卷2期 (2016/06) Pp. 1-13

https://doi.org/10.6966/EDMJ.201606_3(2).0001

摘要

Objectives: Human umbilical cord blood-derived mononuclear cells (UCB-MNCs) have been widely used for the isolation and development of hematopoietic and mesenchymal stem cells (MSCs). This study aimed to establish a research-grade human UCB-MNC bank at E-Da Hospital and to characterize the multipotent differentiation capacity of the isolated cells after ex vivo expansion. Methods: Human UCB-MNCs were isolated from donor mothers using gradient density centrifugation. After ex vivo expansion, adherent cells were tested for MSC surface markers by flow cytometry and their ability to undergo adipogenesis, chondrogenesis, osteogenesis, and hepatogenesis. Results: Despite a low percentage of MNCs from UCB being adherent, the adherent cells grown in MSC expansion medium showed typical mesenchymal morphology and a rapid growth rate. After cryopreservation and thawing, the cells still displayed high viability. No difference was noted between cells stored using either cryopreservation technique. Flow cytometry demonstrated that all isolated adherent UCB-MNCs expressed MSC-characteristic surface antigens, including CD45, CD73, CD90, and CD105, without hematopoietic markers. Moreover, gene expression and phenotypic analyses showed that isolated adherent UCB-MNCs were capable of being induced for adipogenesis, chondrogenesis, osteogenesis, and hepatogenesis. These results support the conclusion that the adherent UCB-MNCs are multipotent MSCs suitable for cell transplantation. Conclusions: We have established a research-grade UCB cell bank with standardized protocols for the isolation and cryopreservation of UCB-MNCs. The multipotency of isolated UCB-MSCs suggests their applicability in further pre-clinical studies and in the development of clinical therapeutic strategies at E-Da Hospital.

關鍵字

adipogenesis ； chondrogenesis ； osteogenesis ； hepatogenesis ； umbilical cord blood-derived mononuclear cells