Production of Proliferation- and Differentiation-Competent Porcine Myoblasts for Preclinical Studies in a Porcine Large Animal Model of Muscular Insufficiency
Muscular insufficiency is observed in many conditions after injury, chronic inflammation, and especially in elderly populations. Causative cell therapies for muscle deficiencies are not state of the art. Animal models to study the therapy efficacy are, therefore, needed. We developed an improved pro...
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MDPI AG
2024-01-01
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| Online Access: | https://www.mdpi.com/2075-1729/14/2/212 |
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| author | Jasmin Knoll Bastian Amend Tanja Abruzzese Niklas Harland Arnulf Stenzl Wilhelm K. Aicher |
| author_facet | Jasmin Knoll Bastian Amend Tanja Abruzzese Niklas Harland Arnulf Stenzl Wilhelm K. Aicher |
| author_sort | Jasmin Knoll |
| collection | DOAJ |
| description | Muscular insufficiency is observed in many conditions after injury, chronic inflammation, and especially in elderly populations. Causative cell therapies for muscle deficiencies are not state of the art. Animal models to study the therapy efficacy are, therefore, needed. We developed an improved protocol to produce myoblasts suitable for pre-clinical muscle therapy studies in a large animal model. Myoblasts were isolated from the striated muscle, expanded by employing five different protocols, and characterized on transcript and protein expression levels to determine procedures that yielded optimized regeneration-competent myoblasts and multi-nucleated myotubes. We report that swine skeletal myoblasts proliferated well under improved conditions without signs of cellular senescence, and expressed significant levels of myogenic markers including Pax7, MyoD1, Myf5, MyoG, Des, Myf6, CD56 (<i>p</i> ≤ 0.05 each). Upon terminal differentiation, myoblasts ceased proliferation and generated multi-nucleated myotubes. Injection of such myoblasts into the urethral sphincter complex of pigs with sphincter muscle insufficiency yielded an enhanced functional regeneration of this muscle (81.54% of initial level) when compared to the spontaneous regeneration in the sham controls without myoblast injection (67.03% of initial level). We conclude that the optimized production of porcine myoblasts yields cells that seem suitable for preclinical studies of cell therapy in a porcine large animal model of muscle insufficiency. |
| first_indexed | 2024-03-07T22:23:54Z |
| format | Article |
| id | doaj.art-8b3fb514552744d9beb076fd566c1e07 |
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| issn | 2075-1729 |
| language | English |
| last_indexed | 2024-03-07T22:23:54Z |
| publishDate | 2024-01-01 |
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| record_format | Article |
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| spelling | doaj.art-8b3fb514552744d9beb076fd566c1e072024-02-23T15:24:39ZengMDPI AGLife2075-17292024-01-0114221210.3390/life14020212Production of Proliferation- and Differentiation-Competent Porcine Myoblasts for Preclinical Studies in a Porcine Large Animal Model of Muscular InsufficiencyJasmin Knoll0Bastian Amend1Tanja Abruzzese2Niklas Harland3Arnulf Stenzl4Wilhelm K. Aicher5Centre of Medical Research, Department of Urology at UKT, Eberhard-Karls-University, 72072 Tuebingen, GermanyDepartment of Urology, University of Tuebingen Hospital, 72076 Tuebingen, GermanyCentre of Medical Research, Department of Urology at UKT, Eberhard-Karls-University, 72072 Tuebingen, GermanyDepartment of Urology, University of Tuebingen Hospital, 72076 Tuebingen, GermanyDepartment of Urology, University of Tuebingen Hospital, 72076 Tuebingen, GermanyCentre of Medical Research, Department of Urology at UKT, Eberhard-Karls-University, 72072 Tuebingen, GermanyMuscular insufficiency is observed in many conditions after injury, chronic inflammation, and especially in elderly populations. Causative cell therapies for muscle deficiencies are not state of the art. Animal models to study the therapy efficacy are, therefore, needed. We developed an improved protocol to produce myoblasts suitable for pre-clinical muscle therapy studies in a large animal model. Myoblasts were isolated from the striated muscle, expanded by employing five different protocols, and characterized on transcript and protein expression levels to determine procedures that yielded optimized regeneration-competent myoblasts and multi-nucleated myotubes. We report that swine skeletal myoblasts proliferated well under improved conditions without signs of cellular senescence, and expressed significant levels of myogenic markers including Pax7, MyoD1, Myf5, MyoG, Des, Myf6, CD56 (<i>p</i> ≤ 0.05 each). Upon terminal differentiation, myoblasts ceased proliferation and generated multi-nucleated myotubes. Injection of such myoblasts into the urethral sphincter complex of pigs with sphincter muscle insufficiency yielded an enhanced functional regeneration of this muscle (81.54% of initial level) when compared to the spontaneous regeneration in the sham controls without myoblast injection (67.03% of initial level). We conclude that the optimized production of porcine myoblasts yields cells that seem suitable for preclinical studies of cell therapy in a porcine large animal model of muscle insufficiency.https://www.mdpi.com/2075-1729/14/2/212cell-based muscle regenerationlarge animal model of cell therapyporcine myoblastmyogenic differentiationmyotubes |
| spellingShingle | Jasmin Knoll Bastian Amend Tanja Abruzzese Niklas Harland Arnulf Stenzl Wilhelm K. Aicher Production of Proliferation- and Differentiation-Competent Porcine Myoblasts for Preclinical Studies in a Porcine Large Animal Model of Muscular Insufficiency Life cell-based muscle regeneration large animal model of cell therapy porcine myoblast myogenic differentiation myotubes |
| title | Production of Proliferation- and Differentiation-Competent Porcine Myoblasts for Preclinical Studies in a Porcine Large Animal Model of Muscular Insufficiency |
| title_full | Production of Proliferation- and Differentiation-Competent Porcine Myoblasts for Preclinical Studies in a Porcine Large Animal Model of Muscular Insufficiency |
| title_fullStr | Production of Proliferation- and Differentiation-Competent Porcine Myoblasts for Preclinical Studies in a Porcine Large Animal Model of Muscular Insufficiency |
| title_full_unstemmed | Production of Proliferation- and Differentiation-Competent Porcine Myoblasts for Preclinical Studies in a Porcine Large Animal Model of Muscular Insufficiency |
| title_short | Production of Proliferation- and Differentiation-Competent Porcine Myoblasts for Preclinical Studies in a Porcine Large Animal Model of Muscular Insufficiency |
| title_sort | production of proliferation and differentiation competent porcine myoblasts for preclinical studies in a porcine large animal model of muscular insufficiency |
| topic | cell-based muscle regeneration large animal model of cell therapy porcine myoblast myogenic differentiation myotubes |
| url | https://www.mdpi.com/2075-1729/14/2/212 |
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