Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts
Abstract Background The pluripotent stem cells in planarians, a model for tissue and cellular regeneration, remain further identification. We recently developed a method to enrich piwi-1+ cells in Schmidtea mediterranea, by staining cells with SiR-DNA and Cell Tracker Green, named SirNeoblasts that...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2021-03-01
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| Series: | Cell Regeneration |
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| Online Access: | https://doi.org/10.1186/s13619-021-00076-6 |
| _version_ | 1818610689607991296 |
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| author | Kaimeng Niu Hao Xu Yuanyi Zhou Xiong Yun Zhao Chong Gao Chris W. Seidel Xue Pan Yuqing Ying Kai Lei |
| author_facet | Kaimeng Niu Hao Xu Yuanyi Zhou Xiong Yun Zhao Chong Gao Chris W. Seidel Xue Pan Yuqing Ying Kai Lei |
| author_sort | Kaimeng Niu |
| collection | DOAJ |
| description | Abstract Background The pluripotent stem cells in planarians, a model for tissue and cellular regeneration, remain further identification. We recently developed a method to enrich piwi-1+ cells in Schmidtea mediterranea, by staining cells with SiR-DNA and Cell Tracker Green, named SirNeoblasts that permits their propagation and subsequent functional study in vivo. Since traditional enrichment for planarian neoblasts by Hoechst 33342 staining generates X1 cells, blocking the cell cycle and inducing cytotoxicity, this method by SiR-DNA and Cell Tracker Green represents a complementary technological advance for functional investigation of cell fate and regeneration. However, the similarities in heterogeneity of cell subtypes between SirNeoblasts and X1 remain unknown. Results In this work, we performed single cell RNA sequencing of SirNeoblasts for comparison with differential expression patterns in a publicly available X1 single cell RNA sequencing data. We found first that all of the lineage-specific progenitor cells in X1 were present in comparable proportions in SirNeoblasts. In addition, SirNeoblasts contain an early muscle progenitor that is unreported in X1. Analysis of new markers for putative pluripotent stem cells identified here, with subsequent sub-clustering analysis, revealed earlier lineages of epidermal, muscular, intestinal, and pharyngeal progenitors than have been observed in X1. Using the gcm as a marker, we also identified a cell subpopulation resided in previously identified tgs-1+ neoblasts. Knockdown of gcm impaired the neoblast repopulation, suggesting a function of gcm in neoblasts. Conclusions In summary, the use of SirNeoblasts will enable broad experimental advances in regeneration and cell fate specification, given the possibility for propagation and transplantation of recombinant and mutagenized pluripotent stem cells that are not previously afforded to this rapid and versatile model system. |
| first_indexed | 2024-12-16T15:18:26Z |
| format | Article |
| id | doaj.art-c5fc91051e3b445e8ca1434547143b24 |
| institution | Directory Open Access Journal |
| issn | 2045-9769 |
| language | English |
| last_indexed | 2024-12-16T15:18:26Z |
| publishDate | 2021-03-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Cell Regeneration |
| spelling | doaj.art-c5fc91051e3b445e8ca1434547143b242022-12-21T22:26:44ZengSpringerOpenCell Regeneration2045-97692021-03-0110111110.1186/s13619-021-00076-6Canonical and early lineage-specific stem cell types identified in planarian SirNeoblastsKaimeng Niu0Hao Xu1Yuanyi Zhou Xiong2Yun Zhao3Chong Gao4Chris W. Seidel5Xue Pan6Yuqing Ying7Kai Lei8Westlake Laboratory of Life Sciences and Biomedicine, Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake UniversityWestlake Laboratory of Life Sciences and Biomedicine, Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake UniversityWestlake Laboratory of Life Sciences and Biomedicine, Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake UniversityWestlake Laboratory of Life Sciences and Biomedicine, Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake UniversityWestlake Laboratory of Life Sciences and Biomedicine, Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake UniversityStowers Institute for Medical ResearchWestlake Laboratory of Life Sciences and Biomedicine, Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake UniversityWestlake Laboratory of Life Sciences and Biomedicine, Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake UniversityWestlake Laboratory of Life Sciences and Biomedicine, Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake UniversityAbstract Background The pluripotent stem cells in planarians, a model for tissue and cellular regeneration, remain further identification. We recently developed a method to enrich piwi-1+ cells in Schmidtea mediterranea, by staining cells with SiR-DNA and Cell Tracker Green, named SirNeoblasts that permits their propagation and subsequent functional study in vivo. Since traditional enrichment for planarian neoblasts by Hoechst 33342 staining generates X1 cells, blocking the cell cycle and inducing cytotoxicity, this method by SiR-DNA and Cell Tracker Green represents a complementary technological advance for functional investigation of cell fate and regeneration. However, the similarities in heterogeneity of cell subtypes between SirNeoblasts and X1 remain unknown. Results In this work, we performed single cell RNA sequencing of SirNeoblasts for comparison with differential expression patterns in a publicly available X1 single cell RNA sequencing data. We found first that all of the lineage-specific progenitor cells in X1 were present in comparable proportions in SirNeoblasts. In addition, SirNeoblasts contain an early muscle progenitor that is unreported in X1. Analysis of new markers for putative pluripotent stem cells identified here, with subsequent sub-clustering analysis, revealed earlier lineages of epidermal, muscular, intestinal, and pharyngeal progenitors than have been observed in X1. Using the gcm as a marker, we also identified a cell subpopulation resided in previously identified tgs-1+ neoblasts. Knockdown of gcm impaired the neoblast repopulation, suggesting a function of gcm in neoblasts. Conclusions In summary, the use of SirNeoblasts will enable broad experimental advances in regeneration and cell fate specification, given the possibility for propagation and transplantation of recombinant and mutagenized pluripotent stem cells that are not previously afforded to this rapid and versatile model system.https://doi.org/10.1186/s13619-021-00076-6PlanariaRegenerationNeoblastSiR-DNAscRNA-seq |
| spellingShingle | Kaimeng Niu Hao Xu Yuanyi Zhou Xiong Yun Zhao Chong Gao Chris W. Seidel Xue Pan Yuqing Ying Kai Lei Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts Cell Regeneration Planaria Regeneration Neoblast SiR-DNA scRNA-seq |
| title | Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts |
| title_full | Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts |
| title_fullStr | Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts |
| title_full_unstemmed | Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts |
| title_short | Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts |
| title_sort | canonical and early lineage specific stem cell types identified in planarian sirneoblasts |
| topic | Planaria Regeneration Neoblast SiR-DNA scRNA-seq |
| url | https://doi.org/10.1186/s13619-021-00076-6 |
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