Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles
Abstract Background Human adipose stromal cells-derived extracellular vesicles (haMSC-EVs) have been shown to alleviate inflammation in acute lung injury (ALI) animal models. However, there are few systemic studies on clinical-grade haMSC-EVs. Our study aimed to investigate the manufacturing, qualit...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-04-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-024-03708-1 |
| _version_ | 1797219988697776128 |
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| author | Jing Wang Zhong-jin Chen Ze-yi Zhang Mei-ping Shen Bo Zhao Wei Zhang Ye Zhang Ji-gang Lei Cheng-jie Ren Jing Chang Cui-li Xu Meng Li Yang-yang Pi Tian-lun Lu Cheng-xiang Dai Su-ke Li Ping Li |
| author_facet | Jing Wang Zhong-jin Chen Ze-yi Zhang Mei-ping Shen Bo Zhao Wei Zhang Ye Zhang Ji-gang Lei Cheng-jie Ren Jing Chang Cui-li Xu Meng Li Yang-yang Pi Tian-lun Lu Cheng-xiang Dai Su-ke Li Ping Li |
| author_sort | Jing Wang |
| collection | DOAJ |
| description | Abstract Background Human adipose stromal cells-derived extracellular vesicles (haMSC-EVs) have been shown to alleviate inflammation in acute lung injury (ALI) animal models. However, there are few systemic studies on clinical-grade haMSC-EVs. Our study aimed to investigate the manufacturing, quality control (QC) and preclinical safety of clinical-grade haMSC-EVs. Methods haMSC-EVs were isolated from the conditioned medium of human adipose MSCs incubated in 2D containers. Purification was performed by PEG precipitation and differential centrifugation. Characterizations were conducted by nanoparticle tracking analysis, transmission electron microscopy (TEM), Western blotting, nanoflow cytometry analysis, and the TNF-α inhibition ratio of macrophage [after stimulated by lipopolysaccharide (LPS)]. RNA-seq and proteomic analysis with liquid chromatography tandem mass spectrometry (LC–MS/MS) were used to inspect the lot-to-lot consistency of the EV products. Repeated toxicity was evaluated in rats after administration using trace liquid endotracheal nebulizers for 28 days, and respiratory toxicity was evaluated 24 h after the first administration. In vivo therapeutic effects were assessed in an LPS-induced ALI/ acute respiratory distress syndrome (ARDS) rat model. Results The quality criteria have been standardized. In a stability study, haMSC-EVs were found to remain stable after 6 months of storage at − 80°C, 3 months at − 20 °C, and 6 h at room temperature. The microRNA profile and proteome of haMSC-EVs demonstrated suitable lot-to-lot consistency, further suggesting the stability of the production processes. Intratracheally administered 1.5 × 108 particles/rat/day for four weeks elicited no significant toxicity in rats. In LPS-induced ALI/ARDS model rats, intratracheally administered haMSC-EVs alleviated lung injury, possibly by reducing the serum level of inflammatory factors. Conclusion haMSC-EVs, as an off-shelf drug, have suitable stability and lot-to-lot consistency. Intratracheally administered haMSC-EVs demonstrated excellent safety at the tested dosages in systematic preclinical toxicity studies. Intratracheally administered haMSC-EVs improved the lung function and exerted anti-inflammatory effects on LPS-induced ALI/ARDS model rats. |
| first_indexed | 2024-04-24T12:42:24Z |
| format | Article |
| id | doaj.art-35557ee36850403693b72a0f7ff77ef1 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-24T12:42:24Z |
| publishDate | 2024-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-35557ee36850403693b72a0f7ff77ef12024-04-07T11:10:30ZengBMCStem Cell Research & Therapy1757-65122024-04-0115112210.1186/s13287-024-03708-1Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesiclesJing Wang0Zhong-jin Chen1Ze-yi Zhang2Mei-ping Shen3Bo Zhao4Wei Zhang5Ye Zhang6Ji-gang Lei7Cheng-jie Ren8Jing Chang9Cui-li Xu10Meng Li11Yang-yang Pi12Tian-lun Lu13Cheng-xiang Dai14Su-ke Li15Ping Li16Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Cellular Biomedicine Group (Shanghai), Co. Ltd.Abstract Background Human adipose stromal cells-derived extracellular vesicles (haMSC-EVs) have been shown to alleviate inflammation in acute lung injury (ALI) animal models. However, there are few systemic studies on clinical-grade haMSC-EVs. Our study aimed to investigate the manufacturing, quality control (QC) and preclinical safety of clinical-grade haMSC-EVs. Methods haMSC-EVs were isolated from the conditioned medium of human adipose MSCs incubated in 2D containers. Purification was performed by PEG precipitation and differential centrifugation. Characterizations were conducted by nanoparticle tracking analysis, transmission electron microscopy (TEM), Western blotting, nanoflow cytometry analysis, and the TNF-α inhibition ratio of macrophage [after stimulated by lipopolysaccharide (LPS)]. RNA-seq and proteomic analysis with liquid chromatography tandem mass spectrometry (LC–MS/MS) were used to inspect the lot-to-lot consistency of the EV products. Repeated toxicity was evaluated in rats after administration using trace liquid endotracheal nebulizers for 28 days, and respiratory toxicity was evaluated 24 h after the first administration. In vivo therapeutic effects were assessed in an LPS-induced ALI/ acute respiratory distress syndrome (ARDS) rat model. Results The quality criteria have been standardized. In a stability study, haMSC-EVs were found to remain stable after 6 months of storage at − 80°C, 3 months at − 20 °C, and 6 h at room temperature. The microRNA profile and proteome of haMSC-EVs demonstrated suitable lot-to-lot consistency, further suggesting the stability of the production processes. Intratracheally administered 1.5 × 108 particles/rat/day for four weeks elicited no significant toxicity in rats. In LPS-induced ALI/ARDS model rats, intratracheally administered haMSC-EVs alleviated lung injury, possibly by reducing the serum level of inflammatory factors. Conclusion haMSC-EVs, as an off-shelf drug, have suitable stability and lot-to-lot consistency. Intratracheally administered haMSC-EVs demonstrated excellent safety at the tested dosages in systematic preclinical toxicity studies. Intratracheally administered haMSC-EVs improved the lung function and exerted anti-inflammatory effects on LPS-induced ALI/ARDS model rats.https://doi.org/10.1186/s13287-024-03708-1Extracellular vesiclesAdipose mesenchymal stromal cellsALI/ARDSInflammation |
| spellingShingle | Jing Wang Zhong-jin Chen Ze-yi Zhang Mei-ping Shen Bo Zhao Wei Zhang Ye Zhang Ji-gang Lei Cheng-jie Ren Jing Chang Cui-li Xu Meng Li Yang-yang Pi Tian-lun Lu Cheng-xiang Dai Su-ke Li Ping Li Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles Stem Cell Research & Therapy Extracellular vesicles Adipose mesenchymal stromal cells ALI/ARDS Inflammation |
| title | Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles |
| title_full | Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles |
| title_fullStr | Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles |
| title_full_unstemmed | Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles |
| title_short | Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles |
| title_sort | manufacturing quality control and glp grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells derived extracellular vesicles |
| topic | Extracellular vesicles Adipose mesenchymal stromal cells ALI/ARDS Inflammation |
| url | https://doi.org/10.1186/s13287-024-03708-1 |
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