Bone marrow stromal cells transduced with a thrombopoietin, interleukin-6, and interleukin-11 syncretic gene induce cord mononuclear cells to generate platelets in vitro.

BACKGROUND

The induction of hematopoietic stem cells to produce mass numbers of platelets (PLTs) in vitro is an effective method to address a lack of PLTs and PLT transfusion resistance in the clinic. However, the design of a low-cost and sustainable culture system is currently problematic.

STUDY DESIGN AND

METHODS

Here, the thrombopoietin, interleukin (IL)-6, and IL-11 genes, three regulatory factors important for thrombopoiesis, were used to construct self-splicing fusion genes linked by foot and mouth disease (F2A) and Theiler's murine encephalitis (T2A) viruses. Bone marrow stromal cells (BMSCs) transduced with the fusion gene acted as nourishing cells and induced cord blood mononuclear cells (MNCs) to generate PLTs in vitro. We counted these cells; determined the percentage of cells expressing specific cell surface markers (CD41); and measured their ability to aggregate via flow cytometry, immunohistochemical staining, and aggregation remote analyzer.

RESULTS

BMSCs transduced with the fusion gene successfully induced cord blood MNCs to generate PLT-sized fragments in the absence of exogenous cytokines. The output was higher than that of the control groups, and the PLT-sized fragments were similar to endogenous PLTs in terms of shape, CD41 expression, and aggregation function.

CONCLUSION

These results suggest that our method could be used to develop a low-cost sustainable cultivation system that generates PLTs in vitro by enhancing the autocrine production of related cytokines through the nourishment provided by cells transduced with a syncretic gene.