A robust and standardized method to isolate and expand mesenchymal stromal cells from human umbilical cord
Cytotherapy, vol. 25, pp. 1057–1068
Abstract
BACKGROUND AIMS: Human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) are increasingly used in research and therapy. To obtain hUC-MSCs, a diversity of isolation and expansion methods are applied. Here, we report on a robust and standardized method for hUC-MSC isolation and expansion. METHODS: Using 90 hUC donors, we compared and optimized critical variables during each phase of the multi-step procedure involving UC collection, processing, MSC isolation, expansion and characterization. Furthermore, we assessed the effect of donor-to-donor variability regarding UC morphology and donor attributes on hUC-MSC characteristics. RESULTS: We demonstrated robustness of our method across 90 UC donors at each step of the procedure. With our method, UCs can be collected up to 6 h after birth, and UC-processing can be initiated up to 48 h after collection without impacting on hUC-MSC characteristics. The removal of blood vessels before explant cultures improved hUC-MSC purity. Expansion in Minimum essential medium α supplemented with human platelet lysate increased reproducibility of the expansion rate and MSC characteristics as compared with Dulbecco's Modified Eagle's Medium supplemented with fetal bovine serum. The isolated hUC-MSCs showed a purity of ∼98.9%, a viability of >97% and a high proliferative capacity. Trilineage differentiation capacity of hUC-MSCs was reduced as compared with bone marrow-derived MSCs. Functional assays indicated that the hUC-MSCs were able to inhibit T-cell proliferation demonstrating their immune-modulatory capacity. CONCLUSIONS: We present a robust and standardized method to isolate and expand hUC-MSCs, minimizing technical variability and thereby lay a foundation to advance reliability and comparability of results obtained from different donors and different studies.
Authors 13
-
Pia Todtenhaupt Aachen Molecular Epidemiology Department of Biomedical Data Sciences Neonatology Department of Pediatrics, Leiden University Medical Center
Leiden University Medical Center · Willem-Alexander Kinderziekenhuis
Affiliation as printed
Molecular Epidemiology, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands; Neonatology, Willem-Alexander Children's Hospital, Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands
Molecular Epidemiology, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands
Neonatology, Willem-Alexander Children's Hospital, Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands
-
Leiden University Medical Center
Affiliation as printed
Molecular Epidemiology, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands
-
Sophie G. Groene Aachen Molecular Epidemiology Department of Biomedical Data Sciences Neonatology Department of Pediatrics, Leiden University Medical Center
Leiden University Medical Center · Willem-Alexander Kinderziekenhuis
Affiliation as printed
Molecular Epidemiology, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands; Neonatology, Willem-Alexander Children's Hospital, Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands
Molecular Epidemiology, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands
Neonatology, Willem-Alexander Children's Hospital, Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands
-
Leiden University Medical Center
Affiliation as printed
Molecular Epidemiology, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands
-
Leiden University Medical Center
Affiliation as printed
Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands; Department of Pathology, Erasmus Medical Center, Leiden, The Netherlands
Department of Pathology, Erasmus Medical Center, Leiden, The Netherlands
Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands
-
Jeanine M.M. van Klink Aachen Neonatology Department of Pediatrics, Leiden University Medical Center
Leiden University Medical Center · Willem-Alexander Kinderziekenhuis
Affiliation as printed
Neonatology, Willem-Alexander Children's Hospital, Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands
-
Arno A.W. Roest Aachen Pediatric Cardiology Department of Pediatrics, Leiden University Medical Center
Leiden University Medical Center · Willem-Alexander Kinderziekenhuis
Affiliation as printed
Pediatric Cardiology, Willem-Alexander Children's Hospital, Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands
-
Christiaan de Bruin Aachen Pediatric Endocrinology Department of Pediatrics, Leiden University Medical Center
Leiden University Medical Center · Willem-Alexander Kinderziekenhuis
Affiliation as printed
Pediatric Endocrinology, Willem-Alexander Children's Hospital, Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands
-
Leiden University Medical Center
Affiliation as printed
Molecular Epidemiology, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands
-
Leiden University Medical Center
Affiliation as printed
Fetal Medicine, Department of Obstetrics, Leiden University Medical Center, Leiden, The Netherlands
-
Leiden University Medical Center · Willem-Alexander Kinderziekenhuis
Affiliation as printed
Neonatology, Willem-Alexander Children's Hospital, Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands
-
Leiden University Medical Center
Affiliation as printed
Molecular Epidemiology, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands
-
Leiden University Medical Center
Affiliation as printed
NecstGen, Leiden, The Netherlands; Department of Internal Medicine, Leiden University Medical Center, Leiden, The Netherlands. Electronic address: m.van_pel@necstgen.com
Department of Internal Medicine, Leiden University Medical Center, Leiden, The Netherlands
NecstGen, Leiden, The Netherlands
Cited by 26 stored of 26
Cited by patents worldwide 1 (Lens.org)
-
TREATMENT OF ORGAN DYSFUNCTION USING A COMBINATION OF MESENCHYMAL STEM CELLS AND EXOSOMESWO2026096838A1 2026-05-07 Pending