Functional Nerve-Vascular Reconstitution of the Bladder-Wall; Application of Patch Transplantation of Skeletal Muscle-Derived Multipotent Stem Cell Sheet-Pellets

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.

Peer Review #2 of "Reconstitution of the complete rupture in musculotendinous junction using skeletal muscle-derived multipotent stem cell sheet-pellets as a “bio-bond” (v0.1)"

“…Here, we used a shorter term expansion culture (in particular, the term of first fiber culture), and this shorter culture period helped preserve the myogenic potential of the sheet-pellets, resulting in new myofiber formation. This was confirmed by the comparison of the RT-PCR data obtained in the previous studies (Soeda et al 2013;Tamaki et al 2013) and the results obtained in this one (Fig. 2).…”

“…5F), because of the role of dystrophin-dystroglycan (α, β) complex in the collagen network (Monti et al 1999;Welser et al 2009). In our previous studies, we prepared the sheet-pellets as the accelerators of the neurovascular reconstitution with diminished myogenic potential Manuscript to be reviewed 20 (Soeda et al 2013;Tamaki et al 2013). Here, we used a shorter term expansion culture (in particular, the term of first fiber culture), and this shorter culture period helped preserve the myogenic potential of the sheet-pellets, resulting in new myofiber formation.…”

“…Single-cell suspension was filtered through 70-, 40-and 20-μm nylon filters in order to remove muscle fibers and other debris, and, after washing, isolated cells were re-cultured in IMDM/20% FCS for 2-3 days, until they reached confluence. In total, cells were expanded in culture for 5-6 days, which represents a reduced culture time compared with previous studies (Soeda et al 2013;Tamaki et al 2013), thus preserving the myogenic potential of sheet-pellets. After they reached confluence, the cells were gently detached from culture dishes using 2 mM EDTA solution.…”

“…Recently, we developed a 3D gel-patch tissue reconstitution system using Sk-MSC sheet-pellets, which are able to preferentially reconstitute peripheral nerve and vascular tissues with robust connective tissue networks (Tamaki et al 2013). Sk-MSC sheet-pellets also expressed various neurotropic/neurotrophic and vasculogenic factor mRNAs before and after transplantation (Soeda et al 2013;Tamaki et al 2013). These properties of Sk-MSCs and their sheet-pellets are considered to be beneficial for the reconstitution of muscle-tendon units, including their nerve-blood vessel networks.…”

Reconstruction of Multiple Facial Nerve Branches Using Skeletal Muscle-Derived Multipotent Stem Cell Sheet-Pellet Transplantation

“…The acceleration of blood vessel formation by Sk-MSCs was also observed with differentiation into vascular endothelial and smooth muscle cells (Figs 4 and 5 ). This ability was also exerted in various tissues [ 10 , 23 – 25 ], and the favorable supply of O 2 /nutrition with the elimination of CO 2 and various waste products in and around the damaged tissue area was expected. Thus, this assisted revascularization may also act as an accelerator of tissue regeneration.…”