A simple, rapid, and efficient method for isolating detrusor for the culture of bladder smooth muscle cells

Ding, Zhi; Xie, Hua; Huang, Yichen; Lv, Yisong; Yang, Gang; Chen, Yan; Sun, Huizhen; Zhou, Junmei; Chen, Fang

doi:10.1007/s11255-015-1138-2

Cited by 3 publications

(4 citation statements)

References 14 publications

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“…The merged image (Fig. 2H) was in accordance with our previous study 18 . Cell viability assay showed that almost all SMCs before harvest were live (Fig.…”

Section: Characterization Of Smc Sheetssupporting

confidence: 89%

Construction of Pedicled Smooth Muscle Tissues by Combining the Capsule Tissue and Cell Sheet Engineering

et al. 2019

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The survival of engineered tissue requires the formation of its own capillary network, which can anastomose with the host vasculature after transplantation. Currently, while many strategies, such as modifying the scaffold material, adding endothelial cells, or angiogenic factors, have been researched, engineered tissue implanted in vivo cannot timely access to sufficient blood supply, leading to ischemic apoptosis or shrinkage. Constructing vascularized engineered tissue with its own axial vessels and subsequent pedicled transfer is promising to solve the problem of vascularization in tissue engineering. In this study, we used the tissue expander capsule as a novel platform for vascularizing autologous smooth muscle cell (SMC) sheets and fabricating vascularized engineered tissue with its own vascular pedicle. First, we verified which time point was the most effective for constructing an axial capsule vascular bed. Second, we compared the outcome of SMC sheet transplantation onto the expander capsule and classical dorsal subcutaneous tissue, which was widely used in other studies for vascularization. Finally, we transplanted multilayered SMC sheets onto the capsule bed twice to verify the feasibility of fabricating thick pedicled engineered smooth muscle tissues. The results indicated that the axial capsule tissue could be successfully induced, and the capsule tissue 1 week after full expansion was the most vascularized. Quantitative comparisons of thickness, vessel density, and apoptosis of cell sheet grafts onto two vascular beds proved that the axial capsule vascular bed was more favorable to the growth and vascularization of transplants than classical subcutaneous tissue. Furthermore, thick vascularized smooth muscle tissues with the vascular pedicle could be constructed by multi-transplanting cell sheets onto the capsule bed. The combination of axial capsule vascular bed and cell sheet engineering may provide an efficient strategy to overcome the problem of slow or insufficient vascularization in tissue engineering.

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“…The merged image (Fig. 2H) was in accordance with our previous study 18 . Cell viability assay showed that almost all SMCs before harvest were live (Fig.…”

Section: Characterization Of Smc Sheetssupporting

confidence: 89%

Construction of Pedicled Smooth Muscle Tissues by Combining the Capsule Tissue and Cell Sheet Engineering

et al. 2019

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“…SMCs and EPCs were cultured as previously described 18 - 20 . Briefly, a pure detrusor strip approximately 10 × 5 mm was cut down from the bladder wall.…”

Section: Methodsmentioning

confidence: 99%

“…Twenty-four rabbits were prepared as described for muscle biopsy 20 , 22 and randomly divided into 3 treatment groups with 8 animals each: 1) cell-sheet transplantation (sheet only group), 2) capsule flap (capsule only group), and 3) vascularized capsule prelaminated with smooth muscle cell sheets (sheet plus capsule group). In each group, the bladder neck (far from the biopsy site) was dissected under a surgical microscope and the muscular layer was separated from the urothelium, followed by a partial detrusorectomy with a mean diameter of 20.0 mm.…”

Section: Methodsmentioning

confidence: 99%

Bladder reconstruction using autologous smooth muscle cell sheets grafted on a pre-vascularized capsule

Guo¹,

Peng²,

Bao³

et al. 2020

Theranostics

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Rationale: Construction of functional vascularized three-dimensional tissues has been a longstanding objective in the field of tissue engineering. The efficacy of using a tissue expander capsule as an induced vascular bed to prefabricate functional vascularized smooth muscle tissue flaps for bladder reconstruction in a rabbit model was tested. Methods: Skin tissue expanders were inserted into the groin to induce vascularized capsule pouch formation. Smooth muscle cells and endothelial progenitor cells were harvested and cocultured to form pre-vascularized smooth muscle cell sheet. Then repeated transplantation of triple-layer cell sheet grafts onto the vascularized capsular tissue was performed at 2-day intervals to prefabricate functional vascularized smooth muscle tissue flaps. Bladder muscular wall defects were created and repaired by six-layer cell sheet graft (sheet only), capsule flap (capsule only) and vascularized capsule prelaminated with smooth muscle cell sheet (sheet plus capsule). The animals were followed for 3 months after implantation and their bladders were explanted serially. Results: Bladder capacity and compliance were maintained in sheet plus capsule group throughout the 3 months. Tissue bath stimulation demonstrated that contractile responses to carbachol and KCl among the three groups revealed a significant difference ( p < 0.05). Histologically, inflammation was evident in the capsule only group at 1 month and fibrosis was observed in sheet only group at 3 months. The vessel density in capsule only and sheet plus capsule group were significantly higher than in the sheet only group at each time point ( p < 0.05). Comparison of the smooth muscle content among the three groups revealed a significant difference ( p < 0.05). Conclusion: These results proved that the capsule may serve as an induced vascular bed for vascularized smooth muscle tissue flap prefabrication. The prefabricated functional vascularized smooth muscle tissue flap has the potential for reliable bladder reconstruction and may create new opportunities for vascularization in 3-D tissue engineering.

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“…Bladder SMCs were isolated as previously described[ 15 ]. Briefly, an approximately 10 × 5 mm smooth muscle strip was excised at vesical vertex and incised into segments.…”

Section: Methodsmentioning

confidence: 99%

Harvesting prevascularized smooth muscle cell sheets from common polystyrene culture dishes

et al. 2018

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Cell sheet engineering has recently emerged as a promising strategy for scaffold-free tissue engineering. However, the primary method of harvesting cell sheets using temperature-responsive dishes has potential limitations. Here we report a novel cell sheet technology based on a coculture system in which SMCs are cocultured with EPCs on common polystyrene dishes. We found that an intact and highly viable cell sheet could be harvested using mechanical methods when SMCs and EPCs were cocultured on common polystyrene dishes at a ratio of 6:1 for 5 to 6 days; the method is simple, cost-effective and highly repeatable. Moreover, the cocultured cell sheet contained capillary-like networks and could secrete a variety of angiogenic factors. Finally, in vivo studies proved that the cocultured cell sheets were more favorable for the fabrication of vascularized smooth muscle tissues compared to single SMC sheets. This study provides a promising avenue for smooth muscle tissue engineering.

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A simple, rapid, and efficient method for isolating detrusor for the culture of bladder smooth muscle cells

Abstract: Pure detrusor can be acquired from bladder wall conveniently using novel method. This novel method brought about significantly higher purity and cell livability as compared to traditional method.

Cited by 3 publications

References 14 publications

Construction of Pedicled Smooth Muscle Tissues by Combining the Capsule Tissue and Cell Sheet Engineering

Construction of Pedicled Smooth Muscle Tissues by Combining the Capsule Tissue and Cell Sheet Engineering

Bladder reconstruction using autologous smooth muscle cell sheets grafted on a pre-vascularized capsule

Harvesting prevascularized smooth muscle cell sheets from common polystyrene culture dishes

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