Optimizing transport methods to preserve function of self‐innervating muscle cells for laryngeal injection
Abstract Objectives Recently, our laboratory has discovered a self‐innervating population of muscle cells, called motor endplate‐expressing cells (MEEs). The cells innately release a wide variety of neurotrophic factors into the microenvironment promoting innervation when used as an injectable treat...
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| Format: | Article |
| Language: | English |
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Wiley
2024-12-01
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| Series: | Laryngoscope Investigative Otolaryngology |
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| Online Access: | https://doi.org/10.1002/lio2.1259 |
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| author | Samuel L. Kaefer Lujuan Zhang Sarah Brookes Rachel A. Morrison Sherry Voytik‐Harbin Stacey Halum |
| author_facet | Samuel L. Kaefer Lujuan Zhang Sarah Brookes Rachel A. Morrison Sherry Voytik‐Harbin Stacey Halum |
| author_sort | Samuel L. Kaefer |
| collection | DOAJ |
| description | Abstract Objectives Recently, our laboratory has discovered a self‐innervating population of muscle cells, called motor endplate‐expressing cells (MEEs). The cells innately release a wide variety of neurotrophic factors into the microenvironment promoting innervation when used as an injectable treatment. Unlike other stem cells, the therapeutic potential of MEEs is dependent on the cells' ability to maintain phenotypical cell surface proteins in particular motor endplates (MEPs). The goal of this study is to identify transport conditions that preserve MEE viability and self‐innervating function. Methods Muscle progenitor cells (MPCs) of adult Yucatan pigs were cultured and induced to generate MEEs. Effects of short‐term cryopreservation methods were studied on MPC and MEE stages. A minimally supplemented medium was investigated for suspension‐mediated transport, and MEEs were loaded at a constant concentration (1 × 107 cells/mL) into plastic syringes. Samples were subjected to varying temperatures (4, 22, and 37°C) and durations (6, 18, 24, and 48 h), which was followed by statistical analysis of viability. Transport conditions maintaining viability acceptable for cellular therapy were examined for apoptosis rates and expression of desired myogenic, neurotrophic, neuromuscular junction, and angiogenic genes. Results Cryopreservation proved detrimental to our cell population. However, a minimally supplemented medium, theoretically safe for injection, was identified. Transport temperature and duration impacted cell viability, with warmer temperatures leading to faster death rates prior to the end of the study. Transport conditions did not appear to affect apoptotic rate. Any expression change of desirable genes fell within the acceptable range. Conclusions Transport state, medium, duration, and temperature must be considered during the transport of injectable muscle cells as they can affect cell viability and expression of integral genes. These described factors are integral in the planning of general cell transport and may prove equally important when the cell population utilized for laryngeal injection is derived from a patient's own initial muscle biopsy. |
| format | Article |
| id | doaj-art-fcdee988d76b4a1eaac21e4a65d884e3 |
| institution | Kabale University |
| issn | 2378-8038 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Laryngoscope Investigative Otolaryngology |
| spelling | doaj-art-fcdee988d76b4a1eaac21e4a65d884e32024-12-23T11:50:36ZengWileyLaryngoscope Investigative Otolaryngology2378-80382024-12-0196n/an/a10.1002/lio2.1259Optimizing transport methods to preserve function of self‐innervating muscle cells for laryngeal injectionSamuel L. Kaefer0Lujuan Zhang1Sarah Brookes2Rachel A. Morrison3Sherry Voytik‐Harbin4Stacey Halum5Indiana University School of Medicine (IUSM) Indianapolis Indiana USADepartment of Otolaryngology‐Head and Neck Surgery IUSM Indianapolis Indiana USAPurdue University Weldon School of Biomedical Engineering West Lafayette Indiana USAPurdue University Weldon School of Biomedical Engineering West Lafayette Indiana USAPurdue University Weldon School of Biomedical Engineering West Lafayette Indiana USAIndiana University School of Medicine (IUSM) Indianapolis Indiana USAAbstract Objectives Recently, our laboratory has discovered a self‐innervating population of muscle cells, called motor endplate‐expressing cells (MEEs). The cells innately release a wide variety of neurotrophic factors into the microenvironment promoting innervation when used as an injectable treatment. Unlike other stem cells, the therapeutic potential of MEEs is dependent on the cells' ability to maintain phenotypical cell surface proteins in particular motor endplates (MEPs). The goal of this study is to identify transport conditions that preserve MEE viability and self‐innervating function. Methods Muscle progenitor cells (MPCs) of adult Yucatan pigs were cultured and induced to generate MEEs. Effects of short‐term cryopreservation methods were studied on MPC and MEE stages. A minimally supplemented medium was investigated for suspension‐mediated transport, and MEEs were loaded at a constant concentration (1 × 107 cells/mL) into plastic syringes. Samples were subjected to varying temperatures (4, 22, and 37°C) and durations (6, 18, 24, and 48 h), which was followed by statistical analysis of viability. Transport conditions maintaining viability acceptable for cellular therapy were examined for apoptosis rates and expression of desired myogenic, neurotrophic, neuromuscular junction, and angiogenic genes. Results Cryopreservation proved detrimental to our cell population. However, a minimally supplemented medium, theoretically safe for injection, was identified. Transport temperature and duration impacted cell viability, with warmer temperatures leading to faster death rates prior to the end of the study. Transport conditions did not appear to affect apoptotic rate. Any expression change of desirable genes fell within the acceptable range. Conclusions Transport state, medium, duration, and temperature must be considered during the transport of injectable muscle cells as they can affect cell viability and expression of integral genes. These described factors are integral in the planning of general cell transport and may prove equally important when the cell population utilized for laryngeal injection is derived from a patient's own initial muscle biopsy.https://doi.org/10.1002/lio2.1259autologous muscle‐derived cellslaryngeal injectionneuro‐muscular junctionneurotrophin |
| spellingShingle | Samuel L. Kaefer Lujuan Zhang Sarah Brookes Rachel A. Morrison Sherry Voytik‐Harbin Stacey Halum Optimizing transport methods to preserve function of self‐innervating muscle cells for laryngeal injection Laryngoscope Investigative Otolaryngology autologous muscle‐derived cells laryngeal injection neuro‐muscular junction neurotrophin |
| title | Optimizing transport methods to preserve function of self‐innervating muscle cells for laryngeal injection |
| title_full | Optimizing transport methods to preserve function of self‐innervating muscle cells for laryngeal injection |
| title_fullStr | Optimizing transport methods to preserve function of self‐innervating muscle cells for laryngeal injection |
| title_full_unstemmed | Optimizing transport methods to preserve function of self‐innervating muscle cells for laryngeal injection |
| title_short | Optimizing transport methods to preserve function of self‐innervating muscle cells for laryngeal injection |
| title_sort | optimizing transport methods to preserve function of self innervating muscle cells for laryngeal injection |
| topic | autologous muscle‐derived cells laryngeal injection neuro‐muscular junction neurotrophin |
| url | https://doi.org/10.1002/lio2.1259 |
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