The Experts below are selected from a list of 18114 Experts worldwide ranked by ideXlab platform
Tetsuro Tamaki - One of the best experts on this subject based on the ideXlab platform.
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reconstitution of the complete rupture in musculotendinous junction using skeletal muscle derived Multipotent Stem Cell sheet pellets as a bio bond
2016Co-Authors: Hiroyuki Hashimoto, Tetsuro Tamaki, Yoshiyasu Uchiyama, Maki Hirata, Masato Sato, Joji MochidaAbstract:Background. Significant and/or complete rupture in the musculotendinous junction (MTJ) is a challenging lesion to treat because of the lack of reliable suture methods. Skeletal muscle-derived Multipotent Stem Cell (Sk-MSC) sheet-pellets, which are able to reconstitute peripheral nerve and muscular/vascular tissues with robust connective tissue networks, have been applied as a “bio-bond”. Methods. Sk-MSC sheet-pellets, derived from GFP transgenic-mice after 7 days of expansion culture, were detached with EDTA to maintain Cell–Cell connections. A completely ruptured MTJ model was prepared in the right tibialis anterior (TA) of the recipient mice, and was covered with sheet-pellets. The left side was preserved as a contralateral control. The control group received the same amount of the Cell-free medium. The sheet-pellet transplantation (SP) group was further divided into two groups; as the short term (4–8 weeks) and long term (14–18 weeks) recovery group. At each time point after transplantation, tetanic tension output was measured through the electrical stimulation of the sciatic nerve. The behavior of engrafted GFP+ tissues and Cells was analyzed by fluorescence immunohistochemistry. Results. The SP short term recovery group showed average 64% recovery of muscle mass, and 36% recovery of tetanic tension output relative to the contralateral side. Then, the SP long term recovery group showed increased recovery of average muscle mass (77%) and tetanic tension output (49%). However, the control group showed no recovery of continuity between muscle and tendon, and demonstrated increased muscle atrophy, with coalescence to the tibia during 4–8 weeks after operation. Histological evidence also supported the above functional recovery of SP group. Engrafted Sk-MSCs primarily formed the connective tissues and muscle fibers, including nerve-vascular networks, and bridged the ruptured tendon–muscle fiber units, with differentiation into skeletal muscle Cells, Schwann Cells, vascular smooth muscle, and endothelial Cells. Discussion. This bridging capacity between tendon and muscle fibers of the Sk-MSC sheet-pellet, as a “bio-bond,” represents a possible treatment for various MTJ ruptures following surgery.
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reconstruction of multiple facial nerve branches using skeletal muscle derived Multipotent Stem Cell sheet pellet transplantation
2015Co-Authors: Kosuke Saito, Tetsuro Tamaki, Maki Hirata, Hiroyuki Hashimoto, Nobuyuki Nakajima, Akihiro Sakai, Kenei Nakazato, Akihito Kazuno, Masahiro Iida, Kenji OkamiAbstract:Head and neck cancer is often diagnosed at advanced stages, and surgical resection with wide margins is generally indicated, despite this treatment being associated with poor postoperative quality of life (QOL). We have previously reported on the therapeutic effects of skeletal muscle-derived Multipotent Stem Cells (Sk-MSCs), which exert reconstitution capacity for muscle-nerve-blood vessel units. Recently, we further developed a 3D patch-transplantation syStem using Sk-MSC sheet-pellets. The aim of this study is the application of the 3D Sk-MSC transplantation syStem to the reconstitution of facial complex nerve-vascular networks after severe damage. Mouse experiments were performed for histological analysis and rats were used for functional examinations. The Sk-MSC sheet-pellets were prepared from GFP-Tg mice and SD rats, and were transplanted into the facial resection model (ST). Culture medium was transplanted as a control (NT). In the mouse experiment, facial-nerve-palsy (FNP) scoring was performed weekly during the recovery period, and immunohistochemistry was used for the evaluation of histological recovery after 8 weeks. In rats, contractility of facial muscles was measured via electrical stimulation of facial nerves root, as the marker of total functional recovery at 8 weeks after transplantation. The ST-group showed significantly higher FNP (about three fold) scores when compared to the NT-group after 2–8 weeks. Similarly, significant functional recovery of whisker movement muscles was confirmed in the ST-group at 8 weeks after transplantation. In addition, engrafted GFP+ Cells formed complex branches of nerve-vascular networks, with differentiation into Schwann Cells and perineurial/endoneurial Cells, as well as vascular endothelial and smooth muscle Cells. Thus, Sk-MSC sheet-pellet transplantation is potentially useful for functional reconstitution therapy of large defects in facial nerve-vascular networks.
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therapeutic isolation and expansion of human skeletal muscle derived Stem Cells for the use of muscle nerve blood vessel reconstitution
2015Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Maki Hirata, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Toshiro Terachi, Joji MochidaAbstract:Skeletal muscle makes up 40-50% of body mass, and is thus considered to be a good adult Stem Cell source for autologous therapy. Although, several Stem/progenitor Cells have been fractionated from mouse skeletal muscle showing a high potential for therapeutic use, it is unclear whether this is the case in human. Differentiation and therapeutic potential of human skeletal muscle-derived Cells (Sk-Cs) was examined. Samples (5-10 g) were obtained from the abdominal and leg muscles of 36 patients (age, 17-79 years) undergoing prostate cancer treatment or leg amputation surgery. All patients gave informed consent. Sk-Cs were isolated using conditioned collagenase solution, and were then sorted as CD34-/CD45-/CD29+ (Sk-DN/29+) and CD34+/CD45- (Sk-34) Cells, in a similar manner as for the previous mouse Sk-Cs. Both Cell fractions were appropriately expanded using conditioned culture medium for about 2 weeks. Differentiation potentials were then examined during Cell culture and in vivo transplantation into the severely damaged muscles of athymic nude mice and rats. Interestingly, these two Cell fractions could be divided into highly myogenic (Sk-DN/29+) and Multipotent Stem Cell (Sk-34) fractions, in contrast to mouse Sk-Cs, which showed comparable capacities in both Cells. At 6 weeks after the separate transplantation of both Cell fractions, the former showed an active contribution to muscle fiber regeneration, but the latter showed vigorous engraftment to the interstitium associated with differentiation into Schwann Cells, perineurial/endoneurial Cells, and vascular endothelial Cells and pericytes, which corresponded to previous observations with mouse SK-Cs. Importantly, mixed cultures of both Cells resulted the reduction of tissue reconstitution capacities in vivo, whereas co-transplantation after separate expansion showed favorable results. Therefore, human Sk-Cs are potentially applicable to therapeutic autografts and show multiple differentiation potential in vivo.
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3d reconstitution of nerve blood vessel networks using skeletal muscle derived Multipotent Stem Cell sheet pellets
2013Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Shuichi Soeda, Akihiro Sakai, Maki Masuda, Nahoko Fukunishi, Toshiro TerachiAbstract:Aim: To cover the large tissue deficits associated with significant loss of function following surgery, a 3D gel-patch-like nerve–vascular reconstitution syStem was developed using the skeletal muscle-derived Multipotent Stem Cell (Sk-MSC) sheet pellet. Materials & methods: The Sk-MSC sheet pellet was prepared from GFP transgenic mice by the collagenase extraction and 7 days expansion Cell culture, and transplanted into a severe muscle damage model with large disruptions to muscle fibers, blood vessels and peripheral nerves. Results: At 4 weeks after transplantation, engrafted Cells contributed to nerve–vascular regeneration associated with Cellular differentiation into Schwann Cells, perineurial/endoneurial Cells, vascular endothelial Cells and pericytes. However, skeletal myogenic differentiation was scarcely observed. Paracrine effects regarding donor Cells/tissues could also be expected, because of the active expression of neurogenic and vasculogenic factor mRNAs in the sheet pellet. Conclusion: These...
Hiroyuki Hashimoto - One of the best experts on this subject based on the ideXlab platform.
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reconstitution of the complete rupture in musculotendinous junction using skeletal muscle derived Multipotent Stem Cell sheet pellets as a bio bond
2016Co-Authors: Hiroyuki Hashimoto, Tetsuro Tamaki, Yoshiyasu Uchiyama, Maki Hirata, Masato Sato, Joji MochidaAbstract:Background. Significant and/or complete rupture in the musculotendinous junction (MTJ) is a challenging lesion to treat because of the lack of reliable suture methods. Skeletal muscle-derived Multipotent Stem Cell (Sk-MSC) sheet-pellets, which are able to reconstitute peripheral nerve and muscular/vascular tissues with robust connective tissue networks, have been applied as a “bio-bond”. Methods. Sk-MSC sheet-pellets, derived from GFP transgenic-mice after 7 days of expansion culture, were detached with EDTA to maintain Cell–Cell connections. A completely ruptured MTJ model was prepared in the right tibialis anterior (TA) of the recipient mice, and was covered with sheet-pellets. The left side was preserved as a contralateral control. The control group received the same amount of the Cell-free medium. The sheet-pellet transplantation (SP) group was further divided into two groups; as the short term (4–8 weeks) and long term (14–18 weeks) recovery group. At each time point after transplantation, tetanic tension output was measured through the electrical stimulation of the sciatic nerve. The behavior of engrafted GFP+ tissues and Cells was analyzed by fluorescence immunohistochemistry. Results. The SP short term recovery group showed average 64% recovery of muscle mass, and 36% recovery of tetanic tension output relative to the contralateral side. Then, the SP long term recovery group showed increased recovery of average muscle mass (77%) and tetanic tension output (49%). However, the control group showed no recovery of continuity between muscle and tendon, and demonstrated increased muscle atrophy, with coalescence to the tibia during 4–8 weeks after operation. Histological evidence also supported the above functional recovery of SP group. Engrafted Sk-MSCs primarily formed the connective tissues and muscle fibers, including nerve-vascular networks, and bridged the ruptured tendon–muscle fiber units, with differentiation into skeletal muscle Cells, Schwann Cells, vascular smooth muscle, and endothelial Cells. Discussion. This bridging capacity between tendon and muscle fibers of the Sk-MSC sheet-pellet, as a “bio-bond,” represents a possible treatment for various MTJ ruptures following surgery.
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reconstruction of multiple facial nerve branches using skeletal muscle derived Multipotent Stem Cell sheet pellet transplantation
2015Co-Authors: Kosuke Saito, Tetsuro Tamaki, Maki Hirata, Hiroyuki Hashimoto, Nobuyuki Nakajima, Akihiro Sakai, Kenei Nakazato, Akihito Kazuno, Masahiro Iida, Kenji OkamiAbstract:Head and neck cancer is often diagnosed at advanced stages, and surgical resection with wide margins is generally indicated, despite this treatment being associated with poor postoperative quality of life (QOL). We have previously reported on the therapeutic effects of skeletal muscle-derived Multipotent Stem Cells (Sk-MSCs), which exert reconstitution capacity for muscle-nerve-blood vessel units. Recently, we further developed a 3D patch-transplantation syStem using Sk-MSC sheet-pellets. The aim of this study is the application of the 3D Sk-MSC transplantation syStem to the reconstitution of facial complex nerve-vascular networks after severe damage. Mouse experiments were performed for histological analysis and rats were used for functional examinations. The Sk-MSC sheet-pellets were prepared from GFP-Tg mice and SD rats, and were transplanted into the facial resection model (ST). Culture medium was transplanted as a control (NT). In the mouse experiment, facial-nerve-palsy (FNP) scoring was performed weekly during the recovery period, and immunohistochemistry was used for the evaluation of histological recovery after 8 weeks. In rats, contractility of facial muscles was measured via electrical stimulation of facial nerves root, as the marker of total functional recovery at 8 weeks after transplantation. The ST-group showed significantly higher FNP (about three fold) scores when compared to the NT-group after 2–8 weeks. Similarly, significant functional recovery of whisker movement muscles was confirmed in the ST-group at 8 weeks after transplantation. In addition, engrafted GFP+ Cells formed complex branches of nerve-vascular networks, with differentiation into Schwann Cells and perineurial/endoneurial Cells, as well as vascular endothelial and smooth muscle Cells. Thus, Sk-MSC sheet-pellet transplantation is potentially useful for functional reconstitution therapy of large defects in facial nerve-vascular networks.
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therapeutic isolation and expansion of human skeletal muscle derived Stem Cells for the use of muscle nerve blood vessel reconstitution
2015Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Maki Hirata, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Toshiro Terachi, Joji MochidaAbstract:Skeletal muscle makes up 40-50% of body mass, and is thus considered to be a good adult Stem Cell source for autologous therapy. Although, several Stem/progenitor Cells have been fractionated from mouse skeletal muscle showing a high potential for therapeutic use, it is unclear whether this is the case in human. Differentiation and therapeutic potential of human skeletal muscle-derived Cells (Sk-Cs) was examined. Samples (5-10 g) were obtained from the abdominal and leg muscles of 36 patients (age, 17-79 years) undergoing prostate cancer treatment or leg amputation surgery. All patients gave informed consent. Sk-Cs were isolated using conditioned collagenase solution, and were then sorted as CD34-/CD45-/CD29+ (Sk-DN/29+) and CD34+/CD45- (Sk-34) Cells, in a similar manner as for the previous mouse Sk-Cs. Both Cell fractions were appropriately expanded using conditioned culture medium for about 2 weeks. Differentiation potentials were then examined during Cell culture and in vivo transplantation into the severely damaged muscles of athymic nude mice and rats. Interestingly, these two Cell fractions could be divided into highly myogenic (Sk-DN/29+) and Multipotent Stem Cell (Sk-34) fractions, in contrast to mouse Sk-Cs, which showed comparable capacities in both Cells. At 6 weeks after the separate transplantation of both Cell fractions, the former showed an active contribution to muscle fiber regeneration, but the latter showed vigorous engraftment to the interstitium associated with differentiation into Schwann Cells, perineurial/endoneurial Cells, and vascular endothelial Cells and pericytes, which corresponded to previous observations with mouse SK-Cs. Importantly, mixed cultures of both Cells resulted the reduction of tissue reconstitution capacities in vivo, whereas co-transplantation after separate expansion showed favorable results. Therefore, human Sk-Cs are potentially applicable to therapeutic autografts and show multiple differentiation potential in vivo.
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3d reconstitution of nerve blood vessel networks using skeletal muscle derived Multipotent Stem Cell sheet pellets
2013Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Shuichi Soeda, Akihiro Sakai, Maki Masuda, Nahoko Fukunishi, Toshiro TerachiAbstract:Aim: To cover the large tissue deficits associated with significant loss of function following surgery, a 3D gel-patch-like nerve–vascular reconstitution syStem was developed using the skeletal muscle-derived Multipotent Stem Cell (Sk-MSC) sheet pellet. Materials & methods: The Sk-MSC sheet pellet was prepared from GFP transgenic mice by the collagenase extraction and 7 days expansion Cell culture, and transplanted into a severe muscle damage model with large disruptions to muscle fibers, blood vessels and peripheral nerves. Results: At 4 weeks after transplantation, engrafted Cells contributed to nerve–vascular regeneration associated with Cellular differentiation into Schwann Cells, perineurial/endoneurial Cells, vascular endothelial Cells and pericytes. However, skeletal myogenic differentiation was scarcely observed. Paracrine effects regarding donor Cells/tissues could also be expected, because of the active expression of neurogenic and vasculogenic factor mRNAs in the sheet pellet. Conclusion: These...
Toshiro Terachi - One of the best experts on this subject based on the ideXlab platform.
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therapeutic isolation and expansion of human skeletal muscle derived Stem Cells for the use of muscle nerve blood vessel reconstitution
2015Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Maki Hirata, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Toshiro Terachi, Joji MochidaAbstract:Skeletal muscle makes up 40-50% of body mass, and is thus considered to be a good adult Stem Cell source for autologous therapy. Although, several Stem/progenitor Cells have been fractionated from mouse skeletal muscle showing a high potential for therapeutic use, it is unclear whether this is the case in human. Differentiation and therapeutic potential of human skeletal muscle-derived Cells (Sk-Cs) was examined. Samples (5-10 g) were obtained from the abdominal and leg muscles of 36 patients (age, 17-79 years) undergoing prostate cancer treatment or leg amputation surgery. All patients gave informed consent. Sk-Cs were isolated using conditioned collagenase solution, and were then sorted as CD34-/CD45-/CD29+ (Sk-DN/29+) and CD34+/CD45- (Sk-34) Cells, in a similar manner as for the previous mouse Sk-Cs. Both Cell fractions were appropriately expanded using conditioned culture medium for about 2 weeks. Differentiation potentials were then examined during Cell culture and in vivo transplantation into the severely damaged muscles of athymic nude mice and rats. Interestingly, these two Cell fractions could be divided into highly myogenic (Sk-DN/29+) and Multipotent Stem Cell (Sk-34) fractions, in contrast to mouse Sk-Cs, which showed comparable capacities in both Cells. At 6 weeks after the separate transplantation of both Cell fractions, the former showed an active contribution to muscle fiber regeneration, but the latter showed vigorous engraftment to the interstitium associated with differentiation into Schwann Cells, perineurial/endoneurial Cells, and vascular endothelial Cells and pericytes, which corresponded to previous observations with mouse SK-Cs. Importantly, mixed cultures of both Cells resulted the reduction of tissue reconstitution capacities in vivo, whereas co-transplantation after separate expansion showed favorable results. Therefore, human Sk-Cs are potentially applicable to therapeutic autografts and show multiple differentiation potential in vivo.
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3d reconstitution of nerve blood vessel networks using skeletal muscle derived Multipotent Stem Cell sheet pellets
2013Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Shuichi Soeda, Akihiro Sakai, Maki Masuda, Nahoko Fukunishi, Toshiro TerachiAbstract:Aim: To cover the large tissue deficits associated with significant loss of function following surgery, a 3D gel-patch-like nerve–vascular reconstitution syStem was developed using the skeletal muscle-derived Multipotent Stem Cell (Sk-MSC) sheet pellet. Materials & methods: The Sk-MSC sheet pellet was prepared from GFP transgenic mice by the collagenase extraction and 7 days expansion Cell culture, and transplanted into a severe muscle damage model with large disruptions to muscle fibers, blood vessels and peripheral nerves. Results: At 4 weeks after transplantation, engrafted Cells contributed to nerve–vascular regeneration associated with Cellular differentiation into Schwann Cells, perineurial/endoneurial Cells, vascular endothelial Cells and pericytes. However, skeletal myogenic differentiation was scarcely observed. Paracrine effects regarding donor Cells/tissues could also be expected, because of the active expression of neurogenic and vasculogenic factor mRNAs in the sheet pellet. Conclusion: These...
Kosuke Saito - One of the best experts on this subject based on the ideXlab platform.
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reconstruction of multiple facial nerve branches using skeletal muscle derived Multipotent Stem Cell sheet pellet transplantation
2015Co-Authors: Kosuke Saito, Tetsuro Tamaki, Maki Hirata, Hiroyuki Hashimoto, Nobuyuki Nakajima, Akihiro Sakai, Kenei Nakazato, Akihito Kazuno, Masahiro Iida, Kenji OkamiAbstract:Head and neck cancer is often diagnosed at advanced stages, and surgical resection with wide margins is generally indicated, despite this treatment being associated with poor postoperative quality of life (QOL). We have previously reported on the therapeutic effects of skeletal muscle-derived Multipotent Stem Cells (Sk-MSCs), which exert reconstitution capacity for muscle-nerve-blood vessel units. Recently, we further developed a 3D patch-transplantation syStem using Sk-MSC sheet-pellets. The aim of this study is the application of the 3D Sk-MSC transplantation syStem to the reconstitution of facial complex nerve-vascular networks after severe damage. Mouse experiments were performed for histological analysis and rats were used for functional examinations. The Sk-MSC sheet-pellets were prepared from GFP-Tg mice and SD rats, and were transplanted into the facial resection model (ST). Culture medium was transplanted as a control (NT). In the mouse experiment, facial-nerve-palsy (FNP) scoring was performed weekly during the recovery period, and immunohistochemistry was used for the evaluation of histological recovery after 8 weeks. In rats, contractility of facial muscles was measured via electrical stimulation of facial nerves root, as the marker of total functional recovery at 8 weeks after transplantation. The ST-group showed significantly higher FNP (about three fold) scores when compared to the NT-group after 2–8 weeks. Similarly, significant functional recovery of whisker movement muscles was confirmed in the ST-group at 8 weeks after transplantation. In addition, engrafted GFP+ Cells formed complex branches of nerve-vascular networks, with differentiation into Schwann Cells and perineurial/endoneurial Cells, as well as vascular endothelial and smooth muscle Cells. Thus, Sk-MSC sheet-pellet transplantation is potentially useful for functional reconstitution therapy of large defects in facial nerve-vascular networks.
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therapeutic isolation and expansion of human skeletal muscle derived Stem Cells for the use of muscle nerve blood vessel reconstitution
2015Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Maki Hirata, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Toshiro Terachi, Joji MochidaAbstract:Skeletal muscle makes up 40-50% of body mass, and is thus considered to be a good adult Stem Cell source for autologous therapy. Although, several Stem/progenitor Cells have been fractionated from mouse skeletal muscle showing a high potential for therapeutic use, it is unclear whether this is the case in human. Differentiation and therapeutic potential of human skeletal muscle-derived Cells (Sk-Cs) was examined. Samples (5-10 g) were obtained from the abdominal and leg muscles of 36 patients (age, 17-79 years) undergoing prostate cancer treatment or leg amputation surgery. All patients gave informed consent. Sk-Cs were isolated using conditioned collagenase solution, and were then sorted as CD34-/CD45-/CD29+ (Sk-DN/29+) and CD34+/CD45- (Sk-34) Cells, in a similar manner as for the previous mouse Sk-Cs. Both Cell fractions were appropriately expanded using conditioned culture medium for about 2 weeks. Differentiation potentials were then examined during Cell culture and in vivo transplantation into the severely damaged muscles of athymic nude mice and rats. Interestingly, these two Cell fractions could be divided into highly myogenic (Sk-DN/29+) and Multipotent Stem Cell (Sk-34) fractions, in contrast to mouse Sk-Cs, which showed comparable capacities in both Cells. At 6 weeks after the separate transplantation of both Cell fractions, the former showed an active contribution to muscle fiber regeneration, but the latter showed vigorous engraftment to the interstitium associated with differentiation into Schwann Cells, perineurial/endoneurial Cells, and vascular endothelial Cells and pericytes, which corresponded to previous observations with mouse SK-Cs. Importantly, mixed cultures of both Cells resulted the reduction of tissue reconstitution capacities in vivo, whereas co-transplantation after separate expansion showed favorable results. Therefore, human Sk-Cs are potentially applicable to therapeutic autografts and show multiple differentiation potential in vivo.
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3d reconstitution of nerve blood vessel networks using skeletal muscle derived Multipotent Stem Cell sheet pellets
2013Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Shuichi Soeda, Akihiro Sakai, Maki Masuda, Nahoko Fukunishi, Toshiro TerachiAbstract:Aim: To cover the large tissue deficits associated with significant loss of function following surgery, a 3D gel-patch-like nerve–vascular reconstitution syStem was developed using the skeletal muscle-derived Multipotent Stem Cell (Sk-MSC) sheet pellet. Materials & methods: The Sk-MSC sheet pellet was prepared from GFP transgenic mice by the collagenase extraction and 7 days expansion Cell culture, and transplanted into a severe muscle damage model with large disruptions to muscle fibers, blood vessels and peripheral nerves. Results: At 4 weeks after transplantation, engrafted Cells contributed to nerve–vascular regeneration associated with Cellular differentiation into Schwann Cells, perineurial/endoneurial Cells, vascular endothelial Cells and pericytes. However, skeletal myogenic differentiation was scarcely observed. Paracrine effects regarding donor Cells/tissues could also be expected, because of the active expression of neurogenic and vasculogenic factor mRNAs in the sheet pellet. Conclusion: These...
Joji Mochida - One of the best experts on this subject based on the ideXlab platform.
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reconstitution of the complete rupture in musculotendinous junction using skeletal muscle derived Multipotent Stem Cell sheet pellets as a bio bond
2016Co-Authors: Hiroyuki Hashimoto, Tetsuro Tamaki, Yoshiyasu Uchiyama, Maki Hirata, Masato Sato, Joji MochidaAbstract:Background. Significant and/or complete rupture in the musculotendinous junction (MTJ) is a challenging lesion to treat because of the lack of reliable suture methods. Skeletal muscle-derived Multipotent Stem Cell (Sk-MSC) sheet-pellets, which are able to reconstitute peripheral nerve and muscular/vascular tissues with robust connective tissue networks, have been applied as a “bio-bond”. Methods. Sk-MSC sheet-pellets, derived from GFP transgenic-mice after 7 days of expansion culture, were detached with EDTA to maintain Cell–Cell connections. A completely ruptured MTJ model was prepared in the right tibialis anterior (TA) of the recipient mice, and was covered with sheet-pellets. The left side was preserved as a contralateral control. The control group received the same amount of the Cell-free medium. The sheet-pellet transplantation (SP) group was further divided into two groups; as the short term (4–8 weeks) and long term (14–18 weeks) recovery group. At each time point after transplantation, tetanic tension output was measured through the electrical stimulation of the sciatic nerve. The behavior of engrafted GFP+ tissues and Cells was analyzed by fluorescence immunohistochemistry. Results. The SP short term recovery group showed average 64% recovery of muscle mass, and 36% recovery of tetanic tension output relative to the contralateral side. Then, the SP long term recovery group showed increased recovery of average muscle mass (77%) and tetanic tension output (49%). However, the control group showed no recovery of continuity between muscle and tendon, and demonstrated increased muscle atrophy, with coalescence to the tibia during 4–8 weeks after operation. Histological evidence also supported the above functional recovery of SP group. Engrafted Sk-MSCs primarily formed the connective tissues and muscle fibers, including nerve-vascular networks, and bridged the ruptured tendon–muscle fiber units, with differentiation into skeletal muscle Cells, Schwann Cells, vascular smooth muscle, and endothelial Cells. Discussion. This bridging capacity between tendon and muscle fibers of the Sk-MSC sheet-pellet, as a “bio-bond,” represents a possible treatment for various MTJ ruptures following surgery.
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therapeutic isolation and expansion of human skeletal muscle derived Stem Cells for the use of muscle nerve blood vessel reconstitution
2015Co-Authors: Tetsuro Tamaki, Yoshiyasu Uchiyama, Maki Hirata, Hiroyuki Hashimoto, Nobuyuki Nakajima, Kosuke Saito, Toshiro Terachi, Joji MochidaAbstract:Skeletal muscle makes up 40-50% of body mass, and is thus considered to be a good adult Stem Cell source for autologous therapy. Although, several Stem/progenitor Cells have been fractionated from mouse skeletal muscle showing a high potential for therapeutic use, it is unclear whether this is the case in human. Differentiation and therapeutic potential of human skeletal muscle-derived Cells (Sk-Cs) was examined. Samples (5-10 g) were obtained from the abdominal and leg muscles of 36 patients (age, 17-79 years) undergoing prostate cancer treatment or leg amputation surgery. All patients gave informed consent. Sk-Cs were isolated using conditioned collagenase solution, and were then sorted as CD34-/CD45-/CD29+ (Sk-DN/29+) and CD34+/CD45- (Sk-34) Cells, in a similar manner as for the previous mouse Sk-Cs. Both Cell fractions were appropriately expanded using conditioned culture medium for about 2 weeks. Differentiation potentials were then examined during Cell culture and in vivo transplantation into the severely damaged muscles of athymic nude mice and rats. Interestingly, these two Cell fractions could be divided into highly myogenic (Sk-DN/29+) and Multipotent Stem Cell (Sk-34) fractions, in contrast to mouse Sk-Cs, which showed comparable capacities in both Cells. At 6 weeks after the separate transplantation of both Cell fractions, the former showed an active contribution to muscle fiber regeneration, but the latter showed vigorous engraftment to the interstitium associated with differentiation into Schwann Cells, perineurial/endoneurial Cells, and vascular endothelial Cells and pericytes, which corresponded to previous observations with mouse SK-Cs. Importantly, mixed cultures of both Cells resulted the reduction of tissue reconstitution capacities in vivo, whereas co-transplantation after separate expansion showed favorable results. Therefore, human Sk-Cs are potentially applicable to therapeutic autografts and show multiple differentiation potential in vivo.
