Introduction: Umbilical cord-derived mesenchymal stem cells, especially from Wharton jelly (WJ-MSCs), are a promising source for regenerative medicine due to their low immunogenicity, immunomodulatory properties, and ethical sourcing. Their integration into biocurative strategies enhances tissue repair. However, clinical applications face challenges such as donor variability, manufacturing inconsistencies, and safety concerns. This study aimed to characterize WJ-MSCs in vitro to support the development of a biocurative product in compliance with Good Manufacturing Practice standards. Methodology: The Wharton jelly from the umbilical cords of four donors was processed using the explant technique for the isolation of WJ-MSCs. They were expanded and characterized by flow cytometry to confirm mesenchymal markers expression. Trilineage differentiation assessed multipotency. Safety evaluations included endotoxin detection (LAL assay), Mycoplasma DNA testing, dimethyl sulfoxide (DMSO) and antibiotic residue analysis (gas and liquid chromatography), and detection of viral particles through culture in Vero 81 and HeLa cells. Results: All samples demonstrated a characteristic mesenchymal immunophenotype, expressing CD73, CD90, and CD105, and lacking expression of CD14, CD34, CD19, and HLA-DR. Multipotency was confirmed by successful trilineage differentiation into adipocytes, chondrocytes, and osteocytes following induction. Endotoxin testing revealed no gel formation, indicating the absence of contamination. All samples tested negative for mycoplasma DNA. No quantifiable levels of DMSO and no residual antibiotics were detected. Additionally, no infectious viral particles were identified in any of the samples. Conclusions: The isolation and expansion protocols were effective. All samples met purity and safety criteria, confirming their suitability for use in advanced therapy medicinal products.
Author Contributions
Conceptualization, M.P.L. and V.M.-S.; methodology, J.C.Z.; software F.C.P.; validation, C.M.C.O.S., V.M.-S. and M.P.L.; formal analysis, J.C.Z.; investigation, D.S.D. and M.P.L.; resources, F.C.P.; data curation, M.P.L., C.M.C.O.S. and F.C.P.; writing—original draft preparation, M.P.L. and J.C.Z.; writing—review and editing, V.M.-S.; supervision, M.P.L.; project administration, M.P.L. and F.C.P.; funding acquisition, V.M.-S. and M.P.L. All authors have read and agreed to the published version of the manuscript.
Funding
This research was funded by Conselho Nacional de Desenvolvimento Científico e Tecnológico–CNPq process n° 440755/2022-0, Curitiba Biotecnologia Ltd and Revolugenix Biotecnologia, Pesquisa e Desenvolvimento Ltd.
Institutional Review Board Statement
The study was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Review Board (or Ethics Committee) of Universidade Positivo (protocol code 6.753.098; CAAE 74617723.4.0000.0093 and approved on 9 April 2024).
Informed Consent Statement
Informed consent was obtained from all subjects involved in the study.
Data Availability Statement
www.curitybabiotech.com.br, accessed on 10 February 2026.
Conflicts of Interest
This research is part of a cell therapy product development project by Curityba Biotech, in partnership with Revolugenix Biotecnologia. Curityba Biotech has no conflicts of interest.
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