TRANSPLANTATION AND CELLULAR ENGINEERING
Novel alginate three-dimensional static and rotating culture systems for effective ex vivo amplification of human cord blood hematopoietic stem cells and in vivo functional analysis of amplified cells in NOD/SCID mice
- This work was supported in part by Grant 81100385 and Grant 81170491 from the National Nature Science Foundation of China.
Address reprint requests to: Yong Xie, PhD, Division of Life Science, Hong Kong University of Science and Technology, Clearwater Bay, Kowloon, Hong Kong SAR, China; e-mail: email@example.com.
Autologous and allogeneic hematopoietic stem cell (HSC) transplantations serve as effective therapy for a variety of hematologic and other diseases. Umbilical cord blood (UCB) is an important source of HSCs. However, it is difficult to obtain a sufficient number of HSCs with complete self-renewal capability derived from a single unit of UCB for use in adult transplantation. In this study, we investigated two novel three-dimensional (3D) culture systems (static and rotating) for ex vivo expansion of HSCs from UCB.
Study Design and Methods
We encapsulated the human cord blood mononuclear cells (CBMCs) in alginate 3D static and rotating culture systems, compared the cell number amplification, the proportion of CD34+ cells, and the colony-forming capacity of these systems to those of the conventional two-dimensional (2D) system. The amplified cells were transplanted into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice to confirm the hematopoiesis reconstruction capacity of the cells.
The increase in the cell number and the proportion of CD34+ cells in the CBMCs was more effective in these 3D alginate culture systems than in the conventional 2D culture system under the same conditions (p < 0.05). The stem cell maintenance capability was confirmed by flow cytometry and colony-forming assay ex vivo and NOD/SCID mice xenogeneic transplantation model in vivo.
Our results demonstrated that these 3D alginate culture systems are an efficient way to amplify cord blood HSCs for extended periods without having them lose their self-renewal capacity in vivo. These novel 3D alginate culture systems are promising for the amplification of UCB-derived HSCs for clinical application in the future.