Academic Journal
Bioorthogonal non-canonical amino acid tagging to track transplanted human induced pluripotent stem cell-specific proteome
| Title: | Bioorthogonal non-canonical amino acid tagging to track transplanted human induced pluripotent stem cell-specific proteome |
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| Authors: | Divya Sridharan, Julie A. Dougherty, Uzair Ahmed, Shridhar K. Sanghvi, Syed Baseeruddin Alvi, Ki Ho Park, Helena Islam, Sue E. Knoblaugh, Harpreet Singh, Elizabeth D. Kirby, Mahmood Khan |
| Source: | Stem Cell Research & Therapy, Vol 15, Iss 1, Pp 1-17 (2024) |
| Publisher Information: | BMC, 2024. |
| Publication Year: | 2024 |
| Collection: | LCC:Medicine (General) LCC:Biochemistry |
| Subject Terms: | Biorthogonal non-canonical amino acid tagging, Human induced pluripotent stem cells, Click chemistry, Cell-specific proteome, Paracrine signaling, Cell transplantation, Medicine (General), R5-920, Biochemistry, QD415-436 |
| More Details: | Abstract Background Human induced pluripotent stem cells (hiPSCs) and their differentiated cell types have a great potential for tissue repair and regeneration. While the primary focus of using hiPSCs has historically been to regenerate damaged tissue, emerging studies have shown a more potent effect of hiPSC-derived paracrine factors on tissue regeneration. However, the precise contents of the transplanted hiPSC-derived cell secretome are ambiguous. This is mainly due to the lack of tools to distinguish cell-specific secretome from host-derived proteins in a complex tissue microenvironment in vivo. Methods In this study, we present the generation and characterization of a novel hiPSC line, L274G-hiPSC, expressing the murine mutant methionyl-tRNA synthetase, L274GMmMetRS, which can be used for tracking the cell specific proteome via biorthogonal non-canonical amino acid tagging (BONCAT). We assessed the trilineage differentiation potential of the L274G-hiPSCs in vitro and in vivo. Furthermore, we assessed the cell-specific proteome labelling in the L274G-hiPSC derived cardiomyocytes (L274G-hiPSC-CMs) in vitro following co-culture with wild type human umbilical vein derived endothelial cells and in vivo post transplantation in murine hearts. Results We demonstrated that the L274G-hiPSCs exhibit typical hiPSC characteristics and that we can efficiently track the cell-specific proteome in their differentiated progenies belonging to the three germ lineages, including L274G-hiPSC-CMs. Finally, we demonstrated cell-specific BONCAT in transplanted L274G-hiPSC-CMs. Conclusion The novel L274G-hiPSC line can be used to study the cell-specific proteome of hiPSCs in vitro and in vivo, to delineate mechanisms underlying hiPSC-based cell therapies for a variety of regenerative medicine applications. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 1757-6512 |
| Relation: | https://doaj.org/toc/1757-6512 |
| DOI: | 10.1186/s13287-024-03792-3 |
| Access URL: | https://doaj.org/article/3817c12ff63f4441b96167eaf8169674 |
| Accession Number: | edsdoj.3817c12ff63f4441b96167eaf8169674 |
| Database: | Directory of Open Access Journals |
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| ISSN: | 17576512 |
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| DOI: | 10.1186/s13287-024-03792-3 |
| Published in: | Stem Cell Research & Therapy |
| Language: | English |