Performance of Developed Mononuclear Cell Separator for Regenerative Therapy Is Comparable to Manual.

Abstract

Regenerative therapy involving transplanted bone marrow mononuclear cells (BM-MNCs) and hematopoietic stem cells (HSCs) is markedly effective against many diseases. However, manual MNC separation requires skilled labor and cell-processing centers. Thus, efforts have been targeted toward fractionating MNCs using cell separators. A double-blind, placebo-controlled study of myocardial infarction using BM-MNCs separated by an existing device was conducted, and no therapeutic effects were found. The development of a cell separator to replace manual techniques would significantly contribute to the widespread application of BM-MNC therapy. Therefore, we developed a BM-MNC separation device that can reproduce manual separation. We changed the shape of the injection port on the centrifuge container and determined its circuit to improve the performance of HSC separation and remove degenerative red blood cells (RBCs). We assessed HSC recovery and degenerative RBC removal rates using fluorescence-activated cell sorting. Additionally, we evaluated the therapeutic effects of cells separated using our device in mouse models of stroke. The HSC recovery and degenerative RBC removal rates using the device were comparable to those obtained using manual separation. We also confirmed the therapeutic effects of BM-MNCs separated by the device in the models. The new automated device could replace manual cell separation and render cell-based therapy using BM-MNC feasible without laborious manual tasks at dedicated cell-processing centers.