Generation of myogenic progenitor cell-derived smooth muscle cells for sphincter regeneration
Abstract Background Degeneration of smooth muscles in sphincters can cause debilitating diseases such as fecal incontinence. Skeletal muscle-derived cells have been effectively used in clinics for the regeneration of the skeletal muscle sphincters, such as the external anal or urinary sphincter. How...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2020-06-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13287-020-01749-w |
| _version_ | 1819228159300599808 |
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| author | Marco Thurner Martin Deutsch Katrin Janke Franka Messner Christina Kreutzer Stanislav Beyl Sébastien Couillard-Després Steffen Hering Jakob Troppmair Rainer Marksteiner |
| author_facet | Marco Thurner Martin Deutsch Katrin Janke Franka Messner Christina Kreutzer Stanislav Beyl Sébastien Couillard-Després Steffen Hering Jakob Troppmair Rainer Marksteiner |
| author_sort | Marco Thurner |
| collection | DOAJ |
| description | Abstract Background Degeneration of smooth muscles in sphincters can cause debilitating diseases such as fecal incontinence. Skeletal muscle-derived cells have been effectively used in clinics for the regeneration of the skeletal muscle sphincters, such as the external anal or urinary sphincter. However, little is known about the in vitro smooth muscle differentiation potential and in vivo regenerative potential of skeletal muscle-derived cells. Methods Myogenic progenitor cells (MPC) were isolated from the skeletal muscle and analyzed by flow cytometry and in vitro differentiation assays. The differentiation of MPC to smooth muscle cells (MPC-SMC) was evaluated by immunofluorescence, flow cytometry, patch-clamp, collagen contraction, and microarray gene expression analysis. In vivo engraftment of MPC-SMC was monitored by transplanting reporter protein-expressing cells into the pyloric sphincter of immunodeficient mice. Results MPC derived from human skeletal muscle expressed mesenchymal surface markers and exhibit skeletal myogenic differentiation potential in vitro. In contrast, they lack hematopoietic surface marker, as well as adipogenic, osteogenic, and chondrogenic differentiation potential in vitro. Cultivation of MPC in smooth muscle differentiation medium significantly increases the fraction of alpha smooth muscle actin (aSMA) and smoothelin-positive cells, while leaving the number of desmin-positive cells unchanged. Smooth muscle-differentiated MPC (MPC-SMC) exhibit increased expression of smooth muscle-related genes, significantly enhanced numbers of CD146- and CD49a-positive cells, and in vitro contractility and express functional Cav and Kv channels. MPC to MPC-SMC differentiation was also accompanied by a reduction in their skeletal muscle differentiation potential. Upon removal of the smooth muscle differentiation medium, a major fraction of MPC-SMC remained positive for aSMA, suggesting the definitive acquisition of their phenotype. Transplantation of murine MPC-SMC into the mouse pyloric sphincter revealed engraftment of MPC-SMC based on aSMA protein expression within the host smooth muscle tissue. Conclusions Our work confirms the ability of MPC to give rise to smooth muscle cells (MPC-SMC) with a well-defined and stable phenotype. Moreover, the engraftment of in vitro-differentiated murine MPC-SMC into the pyloric sphincter in vivo underscores the potential of this cell population as a novel cell therapeutic treatment for smooth muscle regeneration of sphincters. |
| first_indexed | 2024-12-23T10:52:51Z |
| format | Article |
| id | doaj.art-dcc1b4dfa47d4f77bf9bbcc837b7b261 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-23T10:52:51Z |
| publishDate | 2020-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-dcc1b4dfa47d4f77bf9bbcc837b7b2612022-12-21T17:49:51ZengBMCStem Cell Research & Therapy1757-65122020-06-0111111710.1186/s13287-020-01749-wGeneration of myogenic progenitor cell-derived smooth muscle cells for sphincter regenerationMarco Thurner0Martin Deutsch1Katrin Janke2Franka Messner3Christina Kreutzer4Stanislav Beyl5Sébastien Couillard-Després6Steffen Hering7Jakob Troppmair8Rainer Marksteiner9Innovacell Biotechnologie AGInnovacell Biotechnologie AGInnovacell Biotechnologie AGDaniel Swarovski Research Laboratory (DSL), Visceral Transplant and Thoracic Surgery, Medical University of InnsbruckInstitute of Experimental Neuroregeneration, Spinal Cord Injury and Tissue Regeneration Center Salzburg (SCI-TReCS), Paracelsus Medical UniversityDepartment of Pharmacology and Toxicology, University of ViennaInstitute of Experimental Neuroregeneration, Spinal Cord Injury and Tissue Regeneration Center Salzburg (SCI-TReCS), Paracelsus Medical UniversityDepartment of Pharmacology and Toxicology, University of ViennaDaniel Swarovski Research Laboratory (DSL), Visceral Transplant and Thoracic Surgery, Medical University of InnsbruckInnovacell Biotechnologie AGAbstract Background Degeneration of smooth muscles in sphincters can cause debilitating diseases such as fecal incontinence. Skeletal muscle-derived cells have been effectively used in clinics for the regeneration of the skeletal muscle sphincters, such as the external anal or urinary sphincter. However, little is known about the in vitro smooth muscle differentiation potential and in vivo regenerative potential of skeletal muscle-derived cells. Methods Myogenic progenitor cells (MPC) were isolated from the skeletal muscle and analyzed by flow cytometry and in vitro differentiation assays. The differentiation of MPC to smooth muscle cells (MPC-SMC) was evaluated by immunofluorescence, flow cytometry, patch-clamp, collagen contraction, and microarray gene expression analysis. In vivo engraftment of MPC-SMC was monitored by transplanting reporter protein-expressing cells into the pyloric sphincter of immunodeficient mice. Results MPC derived from human skeletal muscle expressed mesenchymal surface markers and exhibit skeletal myogenic differentiation potential in vitro. In contrast, they lack hematopoietic surface marker, as well as adipogenic, osteogenic, and chondrogenic differentiation potential in vitro. Cultivation of MPC in smooth muscle differentiation medium significantly increases the fraction of alpha smooth muscle actin (aSMA) and smoothelin-positive cells, while leaving the number of desmin-positive cells unchanged. Smooth muscle-differentiated MPC (MPC-SMC) exhibit increased expression of smooth muscle-related genes, significantly enhanced numbers of CD146- and CD49a-positive cells, and in vitro contractility and express functional Cav and Kv channels. MPC to MPC-SMC differentiation was also accompanied by a reduction in their skeletal muscle differentiation potential. Upon removal of the smooth muscle differentiation medium, a major fraction of MPC-SMC remained positive for aSMA, suggesting the definitive acquisition of their phenotype. Transplantation of murine MPC-SMC into the mouse pyloric sphincter revealed engraftment of MPC-SMC based on aSMA protein expression within the host smooth muscle tissue. Conclusions Our work confirms the ability of MPC to give rise to smooth muscle cells (MPC-SMC) with a well-defined and stable phenotype. Moreover, the engraftment of in vitro-differentiated murine MPC-SMC into the pyloric sphincter in vivo underscores the potential of this cell population as a novel cell therapeutic treatment for smooth muscle regeneration of sphincters.http://link.springer.com/article/10.1186/s13287-020-01749-wSkeletal muscle-derived cellsSmooth muscleMyogenic progenitor cellsSatellite cellsSmooth muscle regenerationCell therapy |
| spellingShingle | Marco Thurner Martin Deutsch Katrin Janke Franka Messner Christina Kreutzer Stanislav Beyl Sébastien Couillard-Després Steffen Hering Jakob Troppmair Rainer Marksteiner Generation of myogenic progenitor cell-derived smooth muscle cells for sphincter regeneration Stem Cell Research & Therapy Skeletal muscle-derived cells Smooth muscle Myogenic progenitor cells Satellite cells Smooth muscle regeneration Cell therapy |
| title | Generation of myogenic progenitor cell-derived smooth muscle cells for sphincter regeneration |
| title_full | Generation of myogenic progenitor cell-derived smooth muscle cells for sphincter regeneration |
| title_fullStr | Generation of myogenic progenitor cell-derived smooth muscle cells for sphincter regeneration |
| title_full_unstemmed | Generation of myogenic progenitor cell-derived smooth muscle cells for sphincter regeneration |
| title_short | Generation of myogenic progenitor cell-derived smooth muscle cells for sphincter regeneration |
| title_sort | generation of myogenic progenitor cell derived smooth muscle cells for sphincter regeneration |
| topic | Skeletal muscle-derived cells Smooth muscle Myogenic progenitor cells Satellite cells Smooth muscle regeneration Cell therapy |
| url | http://link.springer.com/article/10.1186/s13287-020-01749-w |
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