3D Spheroids of Umbilical Cord Blood MSC-Derived Schwann Cells Promote Peripheral Nerve Regeneration

Abstract

Schwann cells (SCs) are promising candidates for cell therapy due to their ability to promote peripheral nerve regeneration. However, SC-based therapies are hindered by the lack of a clinically renewable source of SCs. In this study, using a well-defined non-genetic approach, umbilical cord blood mesenchymal stem cells (cbMSCs), a clinically applicable cell type, were phenotypically, epigenetically, and functionally converted into SC-like cells (SCLCs) that stimulated effective sprouting of neuritic processes from neuronal cells. To further enhance their therapeutic capability, the cbMSC-derived SCLCs were assembled into three-dimensional (3D) cell spheroids by using a methylcellulose hydrogel system. The cell–cell and cell–extracellular matrix interactions were well-preserved within the formed 3D SCLC spheroids, and marked increases in neurotrophic, proangiogenic and anti-apoptotic factors were detected compared with cells that were harvested using conventional trypsin-based methods, demonstrating the superior advantage of SCLCs assembled into 3D spheroids. Transplantation of 3D SCLC spheroids into crush-injured rat sciatic nerves effectively promoted the recovery of motor function and enhanced nerve structure regeneration. In summary, by simply assembling cells into a 3D-spheroid conformation, the therapeutic potential of SCLCs derived from clinically available cbMSCs for promoting nerve regeneration was enhanced significantly. Thus, these cells hold great potential for translation to clinical applications for treating peripheral nerve injury.

Modulation of Inherent Niches in 3D Multicellular MSC Spheroids Reconfigures Metabolism and Enhances Therapeutic Potential

“…Since the in vivo therapeutic potential of RFP-and TERT-transfected MSCs has been previously verified using various animal models [5][6][7][8], we believe that these MSCs are suitable for use in the present study for further investigation. For a determination of the effect of spheroid size on therapeutic potential, a method that can fabricate spheroids with the desired diameters must be used.…”

“…For a determination of the effect of spheroid size on therapeutic potential, a method that can fabricate spheroids with the desired diameters must be used. Herein, by controlling the number of cells seeded into each well of MC hydrogel-coated plates [5][6][7][8][9], we could precisely manipulate the size of the formed spheroids, which is crucial for ensuring the consistency of the following therapeutic effect after cell transplantation. The harvested MSC spheroids contained abundant matrix components, including fibronectin, laminin, collagen, and HSPG.…”

“…Among the developed methods, we [5][6][7][8][9] and others [2,3,[10][11][12] have reported that culturing MSCs in a three-dimensional (3D) multicellular spheroid configuration can significantly enhance both the viability of administered MSCs and their overall therapeutic functions. Specifically, the spheroid configuration can more closely recapitulate the physiological 3D microenvironment than conventional two-dimensional (2D) cultures [3].…”

“…The MSC spheroids were prepared using a methylcellulose (MC) hydrogel that was not bound by cells. Twelve percent (w/v) MC solution was prepared by dissolving MC powder in phosphate buffered saline (PBS; Thermo Fisher Scientific, St. Louis, MO, USA) [5][6][7][8][9]. After autoclaving, a sterilized MC solution was employed to coat each well of a 96-well culture plate.…”

Chitin Nerve Conduits with Three-Dimensional Spheroids of Mesenchymal Stem Cells from SD Rats Promote Peripheral Nerve Regeneration

“…The qRT-PCR was performed to evaluate the stemness-related gene expression of BMSCs sheets and BMSCs spheroids [20]. Briefly, the total RNA was extracted with the equal cell number of BMSCs sheets and BMSCs spheroids with miRNA kit (Tiangen, Beijing, China).…”