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Sequential passage and characterization of mesenchymal stem cells derived from single and pooled human umbilical cord tissue

Chirayu Padhiar, Muthuraman Muthuchamy, Arvind Kumar Dhanraj, Srividhya Raghavan, Mayur Abhaya, Wilson Aruni.


Human umbilical cord tissue (hUCT) is one of the non-invasive sources of mesenchymal stem cells (MSCs) and comprises similar cell surface markers as that of bone marrow-derived MSCs. Although the MSCs from hUCT are known to have several therapeutic advantages and are found to be implicated in numerous clinical trials, there is often a significant requirement for the expansion of the MSCs at the laboratory level before transplantation. In this regard, the main objective of this study is to compare the variations between cells expanded from single donors and cells pooled from multiple donors. For this, (i) the cells from three individual donors were sequentially expanded up to ten passages (P10) and (ii) the cells from three donors were pooled at P1 and P2 stages, respectively, and expanded sequentially till P10. The expanded cells were characterized for surface markers, trilineage differentiation, and mutational variations at each passage. Furthermore, the cells were pooled and expanded in the 3D stir tank reactor at the P4 passage and checked for variations. Results indicate that the cells at each passage level from both individual and pooled donors were normal concerning the expression of the cell surface markers such as CD90, CD105, CD31, differentiation into osteocyte, chondrocyte, and adipocyte, and no abnormalities were observed in karyotyping. Furthermore, the characteristics of cells expanded in the stirred tank bioreactor were found to be normal. Thus, we conclude that the MSCs can be pooled from the different donors and expanded to the desired numbers before transplantation as there are no variations found at the mutational and characterization levels.

Key words: Umbilical cord, mesenchymal stem cells, stirred tank bioreactor, pooled donors, sequential passaging

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