Abstract
A three-dimensional gel-patch-like nerve-vascular reconstitution system using the Skeletal Muscle-Derived Multipotent Stem Cell (Sk-MSC) sheet-pellet was applied to the reconstitution of the severely damaged bladder wall as a non-skeletal muscle tissue, but has high demand for function. The Sk-MSC sheet-pellet was prepared by the mild detachment of expanded/confluent cells in culture with EDTA, then, collected in a tube and centrifuged. The sheetpellet was pasted on the open thin-walled region of the damaged bladder wall made by myotomy (remove one-third of serosal smooth muscle layer associate with large disruptions of nerve-blood vessel networks retaining the mucosal layer). At 4 wk after transplantation, significant prevention of the reduction in the passive wall-tension, and the positive wall-contraction via electrical stimulation was observed in the transplanted group. Supporting these functional results, immunohistochemical and immunoelectron microscopic analysis revealed that the engrafted cells actively contributed to the reconstitution of blood vessels and peripheral nerves with differentiation into pericytes, endothelial cells, and Schwann cells. However, skeletal and smooth muscle formation was not observed. Thus, this method is potentially useful for the reconstitution of nerve-vascular networks in the bladder-wall to be retaining function such as passive tension and contractile function.
Highlights
The 3D-gel-patch nerve-vascular reconstitution method using expanded Skeletal Muscle-Derived Multipotent Stem Cell (Sk-MSC) sheet-pellets was developed recently [1]
Interstitial cells were divided into the CD34+/CD45− (Sk-34) and CD34−/45− (Sk-DN) fractions at the fresh isolation by collagenase, and both cells were different from satellite cells [5,6,7,8], side-population cells [9] and bone marrow-derived mesenchymal stem cells [2]
The characteristic of this sheet-pellets method is its handling, as the gel-like state allows it to be picked up with forceps, and placed onto the desired site correctly. This state contributes to cell holding and concentration of various paracrine factors in the desired site after transplantation. When this Sk-MSC sheet-pellets was applied for severely damaged skeletal muscle, engrafted cells actively reconstituted nerve-vascular networks associate with cellular differentiation into Schwann cells, perineurial cells with perineurium, endoneurium, vascular endothelial cells and pericytes, whereas a note that, skeletal myogenic differentiation capacity was wholly diminished after expansion culture [1]
Summary
The 3D-gel-patch nerve-vascular reconstitution method using expanded Skeletal Muscle-Derived Multipotent Stem Cell (Sk-MSC) sheet-pellets was developed recently [1]. The characteristic of this sheet-pellets method is its handling, as the gel-like state allows it to be picked up with forceps, and placed onto the desired site correctly This state contributes to cell holding (prevent a cell spread or diffusion) and concentration of various paracrine factors in the desired site after transplantation. When this Sk-MSC sheet-pellets was applied for severely damaged skeletal muscle, engrafted cells actively reconstituted nerve-vascular networks associate with cellular differentiation into Schwann cells, perineurial cells with perineurium, endoneurium, vascular endothelial cells and pericytes, whereas a note that, skeletal myogenic differentiation capacity was wholly diminished after expansion culture [1]. These characteristics strongly suggested its utility as an accelerator and/or adjuvant in other non-skeletal muscle tissue regenerations, requiring nerve-vascular reconstitution
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