| Texto completo | |
| Autor(es): |
Parca, Andressa Valim
;
Pieri, Naira Caroline Godoy
;
Neto, Paulo Fantinato
;
Bressan, Fabiana Fernandes
;
Ambrosio, Carlos Eduardo
;
Martins, Daniele dos Santos
Número total de Autores: 6
|
| Tipo de documento: | Artigo Científico |
| Fonte: | TISSUE ENGINEERING PART C-METHODS; v. 30, n. 4, p. 10-pg., 2024-03-20. |
| Resumo | |
The utmost aim of regenerative medicine is to promote the regeneration of injured tissues using stem cells. Amniotic mesenchymal stem cells (AmMSCs) have been used in several studies mainly because of their easy isolation from amniotic tissue postpartum and immunomodulatory and angiogenic properties and the low level of rejection. These cells share characteristics of both embryonic/fetal and adult stem cells and are particularly advantageous because they do not trigger tumorigenic activity when injected into immunocompromised animals. The large-scale use of AmMSCs for cellular therapies would greatly benefit from fluorescence labeling studies to validate their tracking in future therapies. This study evaluated the fluorophore positivity, fluorescence intensity, and longevity of canine AmMSCs. For this purpose, canine AmMSCs from the GDTI/USP biobank were submitted to three labeling conditions, two commercial fluorophores [CellTrace CFSE Cell Proliferation kit - CTrace, and CellTracker Green CMFDA - CTracker (CellTracker Green CMFDA, CT, #C2925, Molecular Probes (R); Life Technologies)] and green fluorescent protein (GFP) expression after lentiviral transduction, to select the most suitable tracer in terms of adequate persistence and easy handling and analysis that could be used in studies of domestic animals. Fluorescence was detected in all groups; however, the patterns were different. Specifically, CTrace and CTracker fluorescence was detected 6 h after labeling, while GFP was visualized no earlier than 48 h after transduction. Flow cytometry analysis revealed more than 70% of positive cells on day 7 in the CTrace and CTracker groups, while fluorescence decreased significantly to 10% or less on day 20. Variations between repetitions were observed in the GFP group under the present conditions. Our results showed earlier fluorescence detection and more uniform results across repetitions for the commercial fluorophores. In contrast, fluorescence persisted for more extended periods in the GFP group. These results indicate a promising direction for assessing the roles of canine AmMSCs in regenerative medicine without genomic integration. Impact statement Currently, amniotic mesenchymal stem cells (AmMSCs) have been used in study of the treatment of spinal cord injury, or for characterization of their immunomodulator profile in vitro. The large-scale use of AmMSCs for cellular therapies would greatly benefit from fluorescence labeling studies to validate their tracking. In this present study, we evaluated the fluorophore positivity, fluorescence intensity, and longevity of canine AmMSCs in vitro. We observed the fluorescence in all groups of the cells; however, the patterns were different. These results indicate a promising direction for assessing the roles of canine AmMSCs in regenerative medicine without genomic integration. (AU) | |
| Processo FAPESP: | 15/26818-5 - Investigação de mecanismos celulares e moleculares da aquisição da toti- e pluripotência induzida in vitro - modelo translacional |
| Beneficiário: | Fabiana Fernandes Bressan |
| Modalidade de apoio: | Auxílio à Pesquisa - Jovens Pesquisadores |